CN1221309C - Apparatus for corpuscle field and flow separation by sound field force and other active force - Google Patents

Abstract

The invention discloses a device and method for identifying substances by using acoustic field force in field flow separation. The device includes: a reaction pool includes at least one inlet port and at least one outlet port, and the reaction pool has such structural characteristics, That is, when the carrier medium is induced to pass through the reaction cell, the moving speed of the carrier medium at different positions in the reaction cell is different; at least one piezoelectric sensor forms a part of the reaction cell, wherein the pressure The electrical sensor can be excited by at least one electrical signal generated by the electrical signal generator and generate an acoustic wave, resulting in at least one acoustic field force exerted on the substance in the carrier medium, the acoustic field force having a component perpendicular to the direction of movement of the carrier medium. The invention also discloses a method for identifying substances by using acoustic field force in field flow separation.

Description

Device and method for field-flow separation of particles using acoustic field force and other forces

technical field

This invention is related to the field of field flow separation. In particular, this invention discloses a device and method for identifying substances using acoustic field force in field-flow separation, or acoustic field force supplemented by electrophoretic force or dielectrophoretic force.

Background technique

The engineering field is familiar with the methods of electric-field-flow separation (E-FFF) and dielectric-field-flow separation (DEP-FFF). For example, US Patent No. 5,240,618 discloses a method for electric field flow separation, and US Patent Nos. 5,888,370, 5,993,630 and 5,993,632 disclose methods and devices for field flow separation using traditional and expanded dielectrophoresis methods. In electro-field flow separation, the electrophoretic force is used to balance the sedimentation force (applied to large particles, the particle size reaches a few microns or larger) and/or the diffusion force (applied to small particles), and to control the particle velocity distribution in the fluid The equilibrium position (or equilibrium distribution) in . Particles with different charges, sizes, and densities are distributed in different equilibrium positions (or different distribution bands), and pass through the reaction cell at different speeds, so that they can be separated into different parts. In the dielectric-field flow separation method (such as Huang et al, Introducing dielectrophoresis as a new force field for field-flow fractionation, Biophys J.1997 Aug: 73 (2): 1118-29; Markx et al, applications of dielectrics in biotechnology , Trends Biotechnol, 1997 Oct, 15(10): 426-32 and Wang et al, Separation of polystyrene microbeads using dielectric/gravitationalfield-flow-fractionation, Biophys J, 1998 May: 74(5): 2689-701), The vertical component of the dielectrophoretic force is used to balance the settling force and control the equilibrium position of the particles in the fluid. Particles with different dielectric properties are distributed at different heights in the fluid and move at different speeds. Particle mixtures introduced into electro-field flow separation or dielectric-field flow separation cells can be separated into several subclasses according to the time the particles leave the cell. The principle of the electro-field flow separation method was demonstrated for the separation of colloid-adsorbed complexes. The principle of the dielectric-field flow separation method was demonstrated for the separation of synthetic polystyrene beads and biological cells.

However, the existing methods and devices for field flow separation have the following limitations. For electric-field current separation, electrode polarization is a significant problem because most of the applied voltage is lost across the electrode/dielectric interface. Moreover, charge can be used as a parameter of separation only in certain special cases. Similarly, for dielectric-field flow separations, the dielectric properties of particles can only be used as separation parameters in some special cases. And in the dielectric-field flow separation of particles, the forward dielectrophoretic force has never been used. The separation efficiency of existing field flow separation methods is still unsatisfactory in many applications. Thus, it is necessary to further improve the field flow separation method to increase the separation efficiency of the method. It is to this aim that the present invention relates to other aspects of this art.

References involved in the present invention are:

Barmatz MB et al., US patent 4523682, June 1985

Caldwell K.D. and Gao Y-S., Anal.Chem.Vol.65, No.13, July, 1993, 1764-1772.

Lee S. et al, Anal. Chem. Vol.61, p2439-2444, 1989.

Levin S. and Giddings J.C.J.Chem.Tech.Biotechnol., Vol.50, p43-56, 1991.

Giddings, Anal. Chem. Vol. 53, p1170A-1178A, 1981.

Giddings, Science, Vol.260, p1456-1465, 1993.

Huang Y et al., Biophys.J.Vol.73, p1118-1129, 1997.

Huang Y et al., J. Hematoterapy and Stem Cell Research Vol.8, 481-490, 1999

Markx, G.H.et al, J.Liq.Chrom.&Rel.Technol., Vol.20, p2857-2872.1997

Pui P.W.S. et al, Biotechnol, Prog. Vol.11, p146-152, 1995.

Ratanathanawongs S.K. and Giddings J.C. Anal. Chem. Vol.64, p6-15, 1992

Springston et al, Anal.Chem.Vol.59, p344-350, 1987.

Wang et al., Biophys. J. Vol.74, p2689-2701, 1998.

Williams et al., Chem.Eng.Comm.Vol.111, p121-147, 1992.

Williams et al., Chem.Eng.Comm.Vol.130, p143-166, 1994.

Williams et al., Chem. Eng. Sci., Vol.51, p4477-4488, 1996.

Williams et al., Anal. Chem. Vol.69, p349-360, 1997.

Yang J.et al.Anal.Chem.Vol.71, p911-918, 1999

Yasuda K. et al., J.Acoust.Soc.Am.Vol.99(4), p1965-1970, April, 1996 A.

Yasuda K. et al., Jpn.J.Appl.Phys.Vol.35(1), No.5B, p3295-3299.1996 B.

Yasuda K. et al., J.Acoust.Soc.Am., Vol.99(2), p1248-1251, 1996C.

Yasuda K. et al., J.Acoust.Soc.Am.Vol.102(1), p642-645, July, 1997.

Yasuda K. and Kamakura T. Appl. Phys. Lett, Vol.71(13), p1771-1773, Sep.1997.

Yoshioka K. and Kawashima Y. Acustica, Vol.5, pages 167-173, 1955.

Contents of the invention

The present invention discloses methods and required devices for particle identification, manipulation and separation using acoustic field radiation force (or acoustic field force), electrophoretic (E) force, dielectrophoretic (DEP) force, gravity, fluid force and fluid distribution. This new invention and traditional Compared with the methods of field-flow separation, electric-field-flow separation and dielectric-field-flow separation, it has many advantages:

· Particles, such as cells, can be separated according to their own properties, such as size, density, dielectric parameters, charged, etc. Likewise, acoustic impedance is a property of such a particle itself and can be used as a parameter for particle identification and separation.

·Positive dielectrophoretic force can also be used for particle separation—it broadens the field of dielectric field flow separation that can only use negative dielectrophoretic force.

· Better particle separation efficiency can be obtained.

Due to the addition of the acoustic field radiation force (or simply called the acoustic field force) as an additional force, the position of the particles can be affected in the operations of field-flow separation, electric-field-flow separation, and dielectric-field-flow separation, including changing The equilibrium position of the particle, or the distribution zone of the particle includes the equilibrium distribution zone of the particle, so these methods are called acoustic field-field-flow separation, acoustic field-electrophoresis-field-flow separation and acoustic-dielectric-field-flow separation. Here, the particles can be various kinds of substances, such as particulate matter, dissolved substances in a solution, or a combination of these substances. The acoustic field force is induced by acoustic waves (such as standing waves), and the direction of the acoustic field force is perpendicular to the flow direction of the fluid in the reaction cell, and parallel to the direction of the active dielectrophoretic force or electrophoretic force. Standing waves can be generated by various methods, such as using piezoelectric transducers.

The present invention discloses methods and required devices for identifying various types of particles and substances and dissolved substances. Here, recognition includes various meanings, such as separation, identification, differentiation and manipulation of particles. The present invention requires that the particles be suspended in a liquid prior to introduction into the device. The identification process takes place in a device which is a closed thin reaction cell. Particles can be separated in the reaction cell based on differences in their properties, such as density, size, dielectric permittivity, conductivity, surface charge, surface structure, or acoustic impedance. The devices and methods described in the present invention can be used to simultaneously identify different types of particles.

The present invention can be applied to the separation, identification and manipulation of various particles, which can be solid particles such as glass beads, latex particles, liquid particles such as liquid droplets or gaseous particles such as bubbles, etc.; particles can also be organic matter, such as Mammalian cells, bacteria, viruses or other microorganisms can also be inorganic substances, such as metal particles; the shapes of the particles can also vary, such as spherical, elliptical, cubic, pie-shaped and needle-shaped; the size of the particles can also be It can be different, such as gold micelles on the nanometer scale, cells on the micron scale, and microparticle polymers on the millimeter scale. Particles that can be analyzed can be, but are not limited to, biomolecules such as DNA, RNA, chromosomes, protein molecules (such as antibodies), cells, and colloidal particles (such as polystyrene beads).

The invention can be applied to various microparticle separation problems, and is especially suitable for cell separation operations in the field of biomedicine. Examples of particle separations include, but are not limited to, separation of cancer cells from normal cells, separation of metastatic cancer cells from blood cells, separation of fetal nucleated cells from maternal red blood cells/nucleated cells, separation of allogenic cells Virus-infected cells, distinguishing between red and white blood cells, and separating bacteria from human fluid metabolites such as blood and urine, etc. When this invention is used in the field of biological research to analyze living cells, the cells can be separated without modifying the cells with ligands, dyes, antibodies or other methods. In non-biological research areas, too, no modification of the particles to be analyzed is required. Of course, the method provided by this invention is also very suitable for the analysis of modified biological substances. The particle separation process based on this invention has little or no effect on the particles to be analyzed. Living cells can remain unaffected, unmodified, and viable throughout the isolation process. Or it can be said that this separation method has a small enough impact on the cells, and the cells can still be used for further inspection, analysis, identification or growth after separation.

A. Devices using acoustic field force

This invention describes a device for material identification using acoustic field forces in field-flow separation comprising:

a) A reaction tank containing at least one inlet port and one outlet port, and the structural design of the reaction tank should meet the following requirements, that is, when the carrier medium is introduced into the reaction tank, the flow rate of the carrier medium at different positions in the reaction tank is different. Same;

b) At least one piezoelectric sensor constitutes a part of the reaction cell, wherein the piezoelectric sensor can be excited by at least one electrical signal generated by an electrical signal generator and generate sound waves, thereby causing at least one piezoelectric sensor to be applied to the carrier medium An acoustic field force on a medium that has a component perpendicular to the direction of movement of the carrier medium.

A device may include a single inlet port and a single outlet port. Alternatively, multiple ingress and egress ports may also be included. It is best to connect the outlet port directly to a collection or identification device. The outlet port of the reaction pool designed by this invention can have various forms. There can be a single egress port, multiple egress ports, or even an array of multiple egress ports. The width of the outlet port can be the same as that of the reaction cell or only occupy a part of the width of the reaction cell. Particles of different shapes and sizes can be collected from the outlet port. The size of the outlet port can vary from twice the size of the particle to match the width of the reaction cell. In one physical example, the outlet port is formed by one or more tubular structures such as Teflon tubes. These tubular structures can be combined into a composite sample port. Further, the outlet can be directly connected to a fraction collector or a collection hole to collect separated samples. In addition to the collection device, other devices can also be connected to the end of the separation device, such as measurement and diagnostic devices, eg hemocytometers, particle counters and spectrophotometers. Other devices for further examination and analysis of the separated substances may also be connected to the device of the present invention.

The design structure of the reaction cell of the device should enable the fluid (liquid or gas) introduced into the reaction cell to have different flow rates at different positions, and the fluid (liquid or gas) to follow the law of velocity distribution when passing through the reaction cell. For example, a reaction cell may be square in shape, comprising a top, a bottom, and two side walls. The top cover and the bottom surface should be completely parallel or substantially parallel, and the distance between the top cover and the bottom surface is the height of the reaction pool. If the cell contains only one inlet and one outlet, then the distance between the outlet and the inlet is the length of the cell. The two side walls should be perfectly parallel or substantially parallel. The distance between these two parallel side walls is called the width of the reaction cell. The two side walls may be part of a gasket between the top cover and the bottom surface. Washers can be cut into flat hexagonal, oval, circular or other shapes in the middle. In a certain physical example, the reaction pool is a flat structure with a rectangular channel in the middle (that is, the height of the reaction pool is much lower than the length and width of the reaction pool, and the shape of the reaction pool is rectangular). The advantage of the reaction cell with this structure is that when the carrier medium flows through the rectangular channel (or flows through the reaction cell), a parabolic or nearly parabolic velocity distribution can be formed. The flow velocity of the carrier medium close to the top cover and the bottom surface is zero, and the flow velocity increases with the increase of the distance between the carrier medium and the top cover and the bottom surface, reaching a maximum value at half the distance between the top cover and the bottom surface. In some other physical examples, the reaction pool is a long and narrow structure (that is, the width of the reaction pool is much lower than the height and length of the reaction pool, and the shape of the reaction pool is rectangular). The reaction cell can also be circular, elliptical, triangular, rectangular, hexagonal and so on. The reaction cell may consist of a top cover, a bottom surface and a gasket between the top cover and the bottom surface. Washers can be cut with a flat hexagonal hole in the middle, or with oval, round, or other shaped holes. In some physical examples, the reaction pool is a flat structure (ie, the height of the reaction pool is much lower than the length and width of the reaction pool, and the shape of the reaction pool is rectangular). A reaction cell of such a flat configuration has the advantage that a parabolic or approximately parabolic velocity profile can be formed when the carrier medium flows through the reaction cell. The flow velocity of the carrier medium close to the top cover and the bottom surface is zero, and the flow velocity increases with the increase of the distance between the carrier medium and the top cover and the bottom surface, reaching a maximum value at half the distance between the top cover and the bottom surface. In general, for a rectangular channel, the width may be from 1 mm to 20 cm, the height from 20 μm to 10 mm, and the length from 1 cm to 200 cm, with a suitable length of 10 cm to 50 cm. While the above data are given, it is not meant that the invention requires the reaction cells to be of regular geometry. The reaction cell can be made of various materials, such as glass, polymer, plastic, quartz, coated metal, etc., but its structural characteristics need to meet the following requirements: when the carrier medium flows through the reaction cell, different The flow rate of the medium at the location is not the same.

The device includes one or more piezoelectric sensors. Multiple piezoelectric sensors can be excited with the same electrical signal or with different electrical signals. These piezoelectric sensors can be located on the inner or outer surface of the reaction cell, or on a plane substantially parallel to the direction in which the carrier medium flows through the reaction cell.

Usually the amplitude and frequency of the electrical signal output by the electrical signal generator used to excite the piezoelectric sensor can be adjusted.

In an example of a preferred entity, the reaction cell is a tubular structure. One or more piezoelectric transducers are located on the inner or outer surface of the tubular structure.

In another preferred solid example, the reaction cell is formed of a top cover, a bottom surface and two side walls. One or more piezoelectric sensors can be located on the top cover of the reaction cell, or on the bottom of the reaction cell, or on the opposite side of the reaction cell. The height of the reaction cell (the distance between the top cover and the bottom surface) is preferably close to the half wavelength of the standing wave.

The device for identifying substances using acoustic force in field flow separation essentially comprises: a) a reaction cell comprising at least one inlet port and at least one outlet port, said reaction cell having such a structural feature that when a carrier medium is induced through the When the reaction cell is used, the moving speed of the carrier medium at different positions in the reaction cell is not the same; b) at least one piezoelectric sensor is located in a certain part of the reaction cell, wherein the piezoelectric sensor can pass through the electric At least one electrical signal generated by the signal generator excites and generates sound waves, thereby resulting in at least one acoustic field force exerted on the substance in the carrier medium, and the acoustic field force has a component perpendicular to the moving direction of the carrier medium.

B. Devices Using Electrophoresis and Acoustic Field Forces

In another aspect, the invention states that a device for identifying substances using electrophoresis and acoustic field force in field flow separation comprises: a) a reaction cell comprising at least one inlet port and at least one outlet port, said reaction cell having such a structural feature , that is, when the carrier medium is induced to pass through the reaction cell, the moving speed of the carrier medium at different positions in the reaction cell is different; b) at least two electrodes form a part of the reaction cell, and the The electrodes can be excited by at least one electrical signal generated by the electrical signal generator to generate an electric field, thereby causing at least one electrophoretic force on the substance in the carrier medium, the electrophoretic force having a component perpendicular to the direction of movement of the carrier medium. C) At least one piezoelectric sensor constitutes a part of the reaction cell, wherein the piezoelectric sensor can be excited by at least one electrical signal generated by an electrical signal generator to generate sound waves, thereby causing at least one applied to the carrier An acoustic field force on a substance in a medium that has a component perpendicular to the direction of movement of the carrier medium.

The device includes an inlet port and an outlet port, and may include multiple inlet ports and multiple outlet ports. Preferably the outlet port is directly connected to the collection device or identification device. The outlet ports of the reaction pool designed in the present invention can have various forms, such as a single outlet port, multiple outlet ports, or even an array of outlet ports. The width of the outlet port can be the same as that of the reaction cell or only occupy a part of the width of the reaction cell. Particles of different shapes and sizes can be collected from the outlet port. The size of the outlet port can range from twice the particle size to the width of the reaction cell. In one physical example, the outlet port is formed from one or more tubular structures such as Teflon tubes. These tubular structures can be combined into a composite sample port. For example, the outlet port can be connected directly to a fraction collector or to a collection well to collect separated samples. In addition to the collection device, other devices can also be connected to the end of the separation device, such as measurement and diagnostic devices, eg hemocytometers, particle counters and spectrophotometers. Other devices for further examination and analysis of the separated substances may also be connected to the separation device.

The design structure of the reaction pool of the device should enable the fluid (liquid or gas) introduced into the reaction pool to have different flow rates at different positions, and the fluid (liquid or gas) to follow the law of velocity distribution when passing through the reaction pool. For example, the reaction cell can be square in shape, comprising a top, a bottom and two side walls. The top cover and the bottom surface should be completely parallel or substantially parallel, and the distance between the top cover and the bottom surface is the height of the reaction pool. If the cell contains only one inlet port and one outlet port, then the distance between the outlet port and the inlet port is the length of the cell. The two side walls should be perfectly parallel or substantially parallel. The distance between these two parallel side walls is called the width of the reaction cell. The two side walls may be part of a gasket between the top cover and the bottom surface. Washers can have a flat hexagon cut out in the middle, or they can be oval, round, or other shapes. In the physical example, the reaction pool is a flat structure with a rectangular channel in the middle (that is, the height of the reaction pool is much lower than the length and width of the reaction pool, and the shape of the reaction pool is rectangular). The advantage of the reaction cell with this structure is that when the carrier medium flows through the rectangular channel (or flows through the reaction cell), a parabolic or nearly parabolic velocity distribution can be formed. The flow velocity of the carrier medium close to the top cover and the bottom surface is zero, and the flow velocity increases with the increase of the distance between the carrier medium and the top cover and the bottom surface, reaching a maximum value at half the distance between the top cover and the bottom surface. In some other physical examples, the reaction pool is a long and narrow structure (that is, the width of the reaction pool is much lower than the height and length of the reaction pool, and the shape of the reaction pool is rectangular). The reaction cell can also be in the shape of circle, ellipse, triangle, rectangle, hexagon and so on. The reaction cell may consist of a top cover, a bottom surface and a gasket between the top cover and the bottom surface. Washers can be cut into rectangular structures in the middle, or ellipses, circles, or other shapes. In some physical examples, the reaction pool is a flat structure (ie, the height of the reaction pool is much lower than the length and width of the reaction pool, and the shape of the reaction pool is rectangular). A reaction cell of such a flat configuration has the advantage that a parabolic or approximately parabolic velocity profile can be formed when the carrier medium flows through the reaction cell. The flow velocity of the carrier medium close to the top cover and the bottom surface is zero, and the flow velocity increases with the increase of the distance between the carrier medium and the top cover and the bottom surface, reaching a maximum value at half the distance between the top cover and the bottom surface. In general, for rectangular channels, the width may be from 1 mm to 20 cm, the height from 20 μm to 10 mm, and the length from 1 cm to 200 cm, preferably from 10 cm to 50 cm. While the above data are given, it is not meant that the invention requires the reaction cells to be of regular geometry. The reaction cell can be made of various materials, such as glass, polymer, plastic, quartz, coated metal, etc., but its structural characteristics need to meet the following requirements: When the carrier medium flows through the reaction cell, different The flow rate of the medium at the location is not the same.

The device includes two or more electrodes. Each electrode is individually connected to an electrical lead in a bus of electrical leads electrically connected to the electrical signal generator. Electrodes can be arranged on the reaction cell in a horizontal or vertical manner. The electrodes can be arranged in a plane substantially parallel to the moving direction of the carrier medium, preferably in a planar electrode array. The electrode array may be an interdigitated electrode array, an interdigitated castle electrode array, or an interdigitated electrode array with protruding arc-shaped tops. Preferably these electrodes are metal layers coated on the surface of the reaction cell, such as gold layer electrodes. Other metals such as platinum, aluminum, chromium, titanium, copper, and silver may also be used.

The electrical signal generator used to excite the electrodes to generate electrophoretic force can be a DC signal source capable of adjusting voltage amplitude, or an AC signal source capable of adjusting amplitude and frequency. The electrical signal used to excite the electrodes to generate electrophoretic force is preferably a direct current signal or a low frequency alternating current signal.

The device contains one or more piezoelectric sensors. If there are multiple piezoelectric sensors, they can be excited by the same electrical signal or by different electrical signals. A plurality of piezoelectric sensors can be located on the inner or outer surface of the reaction cell, or on a plane substantially parallel to the direction in which the carrier medium flows through the reaction cell.

Generally, the amplitude and frequency of the electric signal output by the electric signal generator for exciting the piezoelectric sensor to generate the acoustic field force can be adjusted.

A common electrical wire bus is used to connect multiple electrodes and signal generators. The common electrical conductor bus can be made in the same way as the electrodes on the device, or it can be one or more conductive elements, such as strip conductors, metal strips or metallized plastic. For interleaved electrode arrays, the interleaved electrodes can be connected together to obtain electrical signals from a signal generator. The signal generator can be a DC power supply that can provide an adjustable voltage. The method and apparatus of the present invention require a voltage in the range of 0 to 15 volts, preferably 0 to 2 volts. The signal generator can also be a power supply that can provide adjustable voltage and frequency, such as Hewlett-Packard's No.8116A signal generator and other multifunctional signal generators. The method and device used in the present invention require a voltage in the range of 0 to 15 volts and a frequency in the range of 0.1 Hz to 100 KHz, and the optimum value is a voltage of 0 to 2 volts and a frequency of 0.1 to 1 KHz.

In an example of a preferred entity, the reaction cell in the device is a tubular structure. Electrodes and/or piezoelectric sensors, or a majority of them, may be located on the inner or outer surface of the tubular structure.

In another example of a preferred entity, the reaction cell consists of a top cover, a bottom surface and two side walls. Electrodes and/or piezoelectric sensors, or most of them can be distributed on the top cover of the reaction cell, or located on the bottom of the reaction cell, in another structure, electrodes and/or piezoelectric sensors, or one of them Most can also be distributed on the opposite side of the reaction tank. The height of the reaction cell (the distance between the top cover and the bottom surface) is preferably close to the half wavelength of the standing wave.

In another example of a preferred entity, the apparatus comprises: a) a reaction well comprising at least one inlet port and at least one outlet port, said reaction well having structural features such that when the carrier medium is induced through the reaction well, The moving speed of the carrier medium at different positions in the reaction cell is not the same; b) at least two electrodes form a part of the reaction cell, wherein the electrodes can pass through at least one path generated by an electrical signal generator electrical signal excitation generates an electric field, resulting in at least one electrophoretic force exerted on the substance in said carrier medium, the electrophoretic force having a component perpendicular to the direction of movement of the carrier medium; C) at least one piezoelectric sensor constituting said reaction cell A part of the method, wherein the piezoelectric sensor can be excited by at least one electrical signal generated by an electrical signal generator to generate acoustic waves, thereby causing at least one acoustic field force applied to the substance in the carrier medium, and the acoustic field force has a force perpendicular to Component of direction of movement of the carrier medium.

C. Devices Using Dielectrophoresis and Acoustic Field Forces

In another aspect, the present invention provides a device for identifying substances using dielectrophoresis and acoustic field force in field flow separation, the device comprising: a) a reaction cell comprising at least one inlet port and at least one outlet port, said reaction cell It has such a structural feature that when the carrier medium is induced to pass through the reaction cell, the moving speed of the carrier medium at different positions in the reaction cell is different; b) at least two electrodes form the reaction cell A part, wherein the electrodes can be excited by at least one electrical signal generated by an electrical signal generator to generate a non-uniform electric field, thereby causing at least one dielectrophoretic force exerted on the substance in the carrier medium, the dielectrophoretic The force has a component perpendicular to the moving direction of the carrier medium; c) at least one piezoelectric sensor forms part of the reaction cell, wherein the piezoelectric sensor can be excited by at least one electrical signal generated by an electrical signal generator The sound waves, thereby causing at least one acoustic field force exerted on the substance in the carrier medium, the acoustic field force having a component perpendicular to the direction of movement of the carrier medium.

The device includes an inlet port and an outlet port, and may include multiple inlet ports and multiple outlet ports. Preferably the outlet port is directly connected to the collection device or identification device. The outlet ports of the reaction pool designed in the present invention can have various forms, such as a single outlet port, multiple outlet ports, or even an array of outlet ports. The width of the outlet port can be the same as that of the reaction cell or only occupy a part of the width of the reaction cell. Particles of different shapes and sizes can be collected from the outlet port. The size of the outlet port can range from twice the particle size to the width of the reaction cell. In one physical example, the outlet port is formed from one or more tubular structures such as Teflon tubes. These tubular structures can be combined into a composite sample port. For example, the outlet port can be connected directly to a fraction collector or to a collection well to collect separated samples. In addition to the collection device, other devices can also be connected to the end of the separation device, such as measurement and diagnostic devices, eg hemocytometers, particle counters and spectrophotometers. Other devices for further examination and analysis of the separated substances may also be connected to the separation device.

The design structure of the reaction pool of the device should enable the fluid (liquid or gas) introduced into the reaction pool to have different flow rates at different positions, and the fluid (liquid or gas) to follow the law of velocity distribution when passing through the reaction pool. For example, the reaction cell can be square in shape, comprising a top, a bottom and two side walls. The top cover and the bottom surface should be completely parallel or substantially parallel, and the distance between the top cover and the bottom surface is the height of the reaction pool. If the cell contains only one inlet port and one outlet port, then the distance between the outlet port and the inlet port is the length of the cell. The two side walls should be perfectly parallel or substantially parallel. The distance between these two parallel side walls is called the width of the reaction cell. The two side walls may be part of a gasket between the top cover and the bottom surface. Washers can have a flat hexagon cut out in the middle, or they can be oval, round, or other shapes. In the physical example, the reaction pool is a flat structure with a rectangular channel in the middle (that is, the height of the reaction pool is much lower than the length and width of the reaction pool, and the shape of the reaction pool is rectangular). The advantage of the reaction cell with this structure is that when the carrier medium flows through the rectangular channel (or flows through the reaction cell), a parabolic or nearly parabolic velocity distribution can be formed. The flow velocity of the carrier medium close to the top cover and the bottom surface is zero, and the flow velocity increases with the increase of the distance between the carrier medium and the top cover and the bottom surface, reaching a maximum value at half the distance between the top cover and the bottom surface. In some other physical examples, the reaction pool is a long and narrow structure (that is, the width of the reaction pool is much lower than the height and length of the reaction pool, and the shape of the reaction pool is rectangular). The reaction cell can also be circular, elliptical, triangular, rectangular, hexagonal and so on. The reaction cell may consist of a top cover, a bottom surface and a gasket between the top cover and the bottom surface. Washers can be cut into rectangular structures in the middle, or they can be cut into ellipses, circles or other shapes. In some physical examples, the reaction pool is a flat structure (ie, the height of the reaction pool is much lower than the length and width of the reaction pool, and the shape of the reaction pool is rectangular). A reaction cell of such a flat configuration has the advantage that a parabolic or approximately parabolic velocity profile can be formed when the carrier medium flows through the reaction cell. The flow velocity of the carrier medium close to the top cover and the bottom surface is zero, and the flow velocity increases with the increase of the distance between the carrier medium and the top cover and the bottom surface, reaching a maximum value at half the distance between the top cover and the bottom surface. In general, for rectangular channels, the width may be from 1 mm to 20 cm, the height from 20 μm to 10 mm, and the length from 1 cm to 200 cm, preferably from 10 cm to 50 cm. While the above data are given, it is not meant that the invention requires the reaction cells to be of regular geometry. The reaction cell can be made of various materials, such as glass, polymer, plastic, quartz, coated metal, etc., but its structural characteristics need to meet the following requirements: When the carrier medium flows through the reaction cell, different The flow rate of the medium at the location is not the same.

The device includes two or more electrodes. Each electrode is individually connected to an electrical lead in a bus of electrical leads electrically connected to the electrical signal generator. Electrodes can be arranged on the reaction cell in a horizontal or vertical manner. The electrodes can also be distributed in a plane substantially parallel to the direction of movement of the carrier medium, preferably arranged in a planar electrode array. The electrode array may be an interdigitated electrode array, an interdigitated castle electrode array, or an interdigitated electrode array with protruding arc-shaped tops. Preferably these electrodes are metal layers coated on the surface of the reaction cell, such as gold layer electrodes. Other metals such as platinum, aluminum, chromium, titanium, copper, and silver may also be used.

The electrical signal generator used to excite the electrodes to generate electrophoretic force can be a DC signal source capable of adjusting voltage amplitude, or an AC signal source capable of adjusting amplitude and frequency.

The device contains one or more piezoelectric sensors. If multiple piezoelectric sensors are suspended, they can be excited by the same electrical signal or by different electrical signals. A plurality of piezoelectric sensors can be located on the inner or outer surface of the reaction cell, or on a plane substantially parallel to the direction in which the carrier medium flows through the reaction cell.

Generally, the amplitude and frequency of the electric signal output by the electric signal generator for exciting the piezoelectric sensor to generate the acoustic field force can be adjusted.

A common electrical wire bus is used to connect multiple electrodes and signal generators. The common electrical conductor bus can be made in the same way as the electrodes on the device, or it can be one or more conductive elements, such as strip conductors, metal strips or metallized plastic. For interleaved electrode arrays, the interleaved electrodes can be connected together to obtain electrical signals from a signal generator. The signal generator can be a DC power supply that can provide an adjustable voltage. The method and apparatus of the present invention require a voltage in the range of 0 to 15 volts, preferably 0 to 10 volts. The signal generator can also be a power supply that can provide adjustable voltage and frequency, such as Hewlett-Packard's No.8116A signal generator and other multifunctional signal generators. The method and apparatus used in the present invention require a voltage in the range of 0 to 15 volts and a frequency in the range of 0.1 Hz to 500 MHz, and the optimum value is a voltage of 0 to 10 volts and a frequency of 0.1 to 10 MHz. Since the frequency required to identify substances using dielectrophoretic forces depends on the properties of the substance to be identified and the conductivity of the medium (eg, cell suspension), the frequencies described here are merely examples. The recognition of substances also depends on the shape, size and arrangement of electrodes. In one practical example, the signal used is a sine wave, although other periodic or non-periodic waveforms may of course be used. The electrical signal may be generated by one or more electrical signal generators capable of adjusting the amplitude and frequency of the output electrical signal.

In an example of a preferred entity, the reaction cell in the device is a tubular structure. Electrodes and/or piezoelectric sensors, or a majority of them, may be located on the inner or outer surface of the tubular structure.

In another example of a preferred entity, the reaction cell consists of a top cover, a bottom surface and two side walls. Electrodes and/or piezoelectric sensors, or most of them can be distributed on the top cover of the reaction cell, or located on the bottom of the reaction cell, in another structure, electrodes and/or piezoelectric sensors, or one of them Most can also be distributed on the opposite side of the reaction tank. The height of the reaction cell (the distance between the top cover and the bottom surface) is preferably close to the half wavelength of the standing wave.

In another example of a preferred entity, the apparatus comprises: a) a reaction well comprising at least one inlet port and at least one outlet port, said reaction well having structural features such that when the carrier medium is induced through the reaction well, The moving speed of the carrier medium at different positions in the reaction cell is not the same; b) at least two electrodes form a part of the reaction cell, wherein the electrodes can pass through at least one path generated by an electrical signal generator electrical signal excitation generates an electric field, resulting in at least one electrophoretic force exerted on the substance in said carrier medium, the electrophoretic force having a component perpendicular to the direction of movement of the carrier medium; C) at least one piezoelectric sensor constituting said reaction cell A part of the method, wherein the piezoelectric sensor can be excited by at least one electrical signal generated by an electrical signal generator to generate acoustic waves, thereby causing at least one acoustic field force applied to the substance in the carrier medium, and the acoustic field force has a force perpendicular to Component of direction of movement of the carrier medium.

D. Methods of identifying substances using acoustic field forces or using acoustic field forces simultaneously with electrophoretic or dielectrophoretic forces

Viewed from another aspect, the invention provides a "continuous mode" method of using acoustic field forces to identify matter in field-flow separation, the method comprising: a) obtaining a device as described in Section A; b) by said device The inlet port of the inlet port will introduce the carrier medium containing the substance to be identified into the reaction cell of the device, wherein the introduction makes the carrier medium pass through the reaction cell according to the velocity distribution; c) by applying at least Another electrical signal excites and generates sound waves, thereby causing at least one acoustic field force applied to the substance in the carrier medium to have a component perpendicular to the direction in which the carrier medium moves through the reaction cell, and the The substance is displaced to a certain position in a direction perpendicular to the moving direction of the carrier medium under the action of force, and the substance is identified according to the difference in the distribution position of the substance in this direction. This is the continuous mode of soundfield-field-flow separation (soundfield-FFF). It can use any of the devices described in Section A.

In another aspect, the present invention provides a "batch mode" method for identifying substances using acoustic field forces in field flow separation. This method comprises: a) obtaining a device as described in section A; b) introducing carrier medium into the reaction cell of the device through the inlet port of the device until the carrier medium fills the entire reaction cell; The sample of the substance is transported (such as injected) into the carrier medium in the reaction cell; d) applying at least one electrical signal generated by the electrical signal generator to the piezoelectric sensor, wherein the excitation of the piezoelectric sensor generates sound waves, resulting in at least There is an acoustic field force exerted on the substance; e) introducing the carrier medium into the reaction cell of the device through the inlet port of the device, wherein the introduction causes the carrier medium to pass through the reaction cell according to the velocity distribution, and the carrier medium in the The substance described in is displaced to a certain position in a direction perpendicular to the moving direction of the carrier medium under the action of force, and the substance is identified according to the difference in the distribution position of the substance in this direction. This "batch mode" of field-field flow separation can use any of the devices described in Section A.

In the "batch mode" method of acoustic field-field flow separation described above, the best method is that it passes the carrier medium through the reaction cell according to a velocity profile (step e) before guiding the carrier medium into the reaction cell (step e), on the piezoelectric transducer The application of an electrical signal causes an acoustic field force to act on the substance which causes the substance to be distributed in an equilibrium position along a direction perpendicular to the flow of the medium through the reaction cell.

In another aspect, the present invention provides a "continuous mode" method for identifying substances using electrophoretic and acoustic field forces simultaneously in field flow separation. This method comprises: a) obtaining a device as described in section B; b) introducing a carrier medium containing a substance to be identified into a reaction cell of the device through an inlet port of the device, wherein said introduction causes the carrier medium passing through the reaction cell according to the velocity distribution; c) generating an electric field by applying at least one electric signal generated by the electric signal generator to the electrode, thereby causing at least one electrophoretic force applied to the substance in the carrier medium to have a direction perpendicular to the carrier The medium moves through the component of the direction of the reaction cell; d) by applying at least one other electrical signal generated by the electrical signal generator to the piezoelectric sensor to excite and generate sound waves, thereby causing at least one acoustic field force applied to the substance in the carrier medium has a component perpendicular to the direction in which the carrier medium moves through the reaction cell, the substance in the carrier medium is displaced to a position in a direction perpendicular to the direction in which the carrier medium moves under the action of a force, according to The difference in the distribution position of the substance in this direction is used to identify the substance. Any of the devices described in Section B can be used in a continuous manner for this acoustic field-electrophoretic-field-flow separation (acoustic-electrophoretic-field-flow separation).

In another aspect, the present invention provides a "batch mode" method for identifying substances using both electrophoretic and acoustic force in field flow separation. This method includes: a) obtaining a device as described in Section B; b) introducing carrier medium into the reaction cell of the device through the inlet port of the device until the carrier medium fills the entire reaction cell; The sample of the substance is transported (such as injected) into the carrier medium in the reaction cell; d) applying at least one electrical signal generated by the electrical signal generator to the electrode, wherein the excitation electrode generates an electric field, resulting in at least one electrophoresis a force is applied to said substance; e) applying at least another electrical signal generated by an electrical signal generator to the piezoelectric sensor, wherein said excitation of the piezoelectric sensor generates an acoustic wave, thereby causing at least one acoustic field force to be applied to the piezoelectric sensor On the substance; f) introducing the carrier medium into the reaction cell of the device through the inlet port of the device, wherein the introduction makes the carrier medium pass through the reaction cell according to the velocity distribution, and the substance in the carrier medium Under the action of force, it is displaced to a certain position in a direction perpendicular to the moving direction of the carrier medium, and the substance is identified according to the difference in the distribution position of the substance in this direction. Any of the devices described in Section B can be used in "batch mode" for this acoustic-electrophoretic-field-flow separation.

In the "batch mode" method of acoustic field-electrophoresis-field flow separation described above, the best method is that it causes the carrier medium to pass through the reaction cell according to a velocity profile (step f) before introducing the carrier medium into the reaction cell (step f). Applying an electrical signal to the sensor produces an acoustic field force acting on the substance, which causes the substance to move to an equilibrium position in a direction perpendicular to the flow of the medium through the reaction cell.

In another aspect, the present invention provides a "continuous mode" method of simultaneously using dielectrophoretic and acoustic field forces to identify substances in field-flow separations, the method comprising: a) obtaining a device as described in Section C; b) introducing the carrier medium into the reaction cell of the device through the inlet port of the device, wherein the introduction causes the carrier medium to pass through the reaction cell according to the velocity distribution; c) applying at least one electrical signal generated by an electrical signal generator to the electrodes, Wherein said excitation electrode generates a non-uniform electric field, thereby causing at least one dielectrophoretic force exerted on said substance to have a component perpendicular to the direction in which the carrier medium moves through the reaction cell; d) for piezoelectric sensors applying at least one other electrical signal generated by an electrical signal generator, wherein said excitation of the piezoelectric sensor generates an acoustic wave, thereby causing at least one acoustic field force on said substance to have a movement perpendicular to the carrier medium through the reaction The component of the pool direction; the substance in the carrier medium is displaced to a certain position in a direction perpendicular to the moving direction of the carrier medium under the action of force, according to the difference in the distribution position of the substance in this direction to identify the substance. Any of the devices described in Section C can be used in the present acoustic field-dielectrophoresis-field-flow separation (acoustic-dielectric-field-flow separation) continuous manner.

In another aspect, the present invention provides a "batch mode" method of simultaneously using dielectrophoretic and acoustic field forces to identify substances in field flow separation, the method comprising: a) obtaining a device as described in section C; b) The carrier medium is introduced into the reaction pool of the device through the inlet port of the device until the carrier medium fills the entire reaction pool; c) the sample containing the substance to be identified is transported (such as injected) into the carrier medium in the reaction pool; d) The electrodes apply at least one electrical signal generated by the electrical signal generator, wherein the excitation electrode generates an electric field, thereby causing at least one dielectrophoretic force to be applied to the substance; e) applying the piezoelectric sensor by at least one other electrical signal generated by an electrical signal generator, wherein said excitation of the piezoelectric sensor generates an acoustic wave, thereby causing at least one acoustic field force to be exerted on said substance; f) introducing the carrier medium through the inlet port of the device Into the reaction pool of the device, wherein the introduction causes the carrier medium to pass through the reaction pool according to the velocity distribution; the substance in the carrier medium is displaced to a direction perpendicular to the moving direction of the carrier medium under the action of force A certain position on the direction, the substance is identified according to the difference in the distribution position of the substance in this direction. Any of the devices described in Section B can be used in a "batch mode" of this acoustic-dielectric-field flow separation.

In the "batch mode" method of acoustic-dielectric-field flow separation described above, it is best to pass the carrier medium through the reaction cell according to a velocity profile (step f) before introducing the carrier medium into the reaction cell (step f), where the piezoelectric The application of an electrical signal to the sensor generates an acoustic field force on the substance which causes the substance to be distributed in an equilibrium position along a direction perpendicular to the flow of the medium through the reaction cell.

In the acousto-electrophoretic-field-flow separation or acousto-dielectric-field-flow separation methods described above, similar but preferably different electrical signals are used to generate the acoustic, electrophoretic or dielectrophoretic forces.

In the above acoustic field-electrophoretic-field-flow separation or acoustic-dielectric-field-flow separation method, the acoustic field force, electrophoretic force or dielectrophoretic force can be generated simultaneously or sequentially.

The above-mentioned acoustic field-electrophoresis-field flow separation, acoustic field-electrophoresis-field flow separation and acoustic field-dielectric-field flow separation method can further include a velocity distribution according to the carrier medium passing through the reaction cell and the material in the reaction cell in the velocity distribution The position determines the speed at which the material moves to identify the steps.

The acoustic field-electrophoretic-field-flow separation, acoustic-electrophoretic-field-flow separation and acoustic-dielectric-field-flow separation methods described above may further comprise a step of separating the identified substance from the device, and preferably results in a separation between the identified substance are also separated from each other. After separation in the carrier medium of the reaction cell according to the invention, the separated substances exit the single or multiple outlet ports of the reaction cell at different times depending on the position of the species in the velocity profile of the carrier medium as it passes through the reaction cell. In particular, substances distributed at different heights in the velocity distribution flow at different velocities, so different substances can be identified by different positions and moving velocities in the velocity distribution. Species at different locations in the velocity profile move at different velocities and exit the single or multiple outlet ports of the reaction cell at different times.

The velocity profile may be a dynamic fluid profile such as a parabolic flow profile. For a rectangular reaction cell, the structural characteristics of the reaction cell can be defined by the length of the reaction cell, the width of the reaction cell, and the height of the reaction cell. The velocity profile can be determined from the known fluid flow velocity, and the width, thickness and length of the reaction cell. For example, for a rectangular reaction cell whose length and width are much greater than the height of the reaction cell, a thin layer distribution can be established in the reaction cell. The velocity of the carrier medium at different positions is mainly determined by the distance from it to the bottom wall of the reaction tank and the velocity distribution approximates the parabolic fluid distribution (or approximates the parabolic distribution), given by

${\mathrm{<span\; class="notranslate">\; V</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}<span\; class="notranslate">\; =</span>\mathrm{<span\; class="notranslate">\; 6</span>}<span\; class="notranslate">\; <</span>{\mathrm{<span\; class="notranslate">\; V</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}<span\; class="notranslate">\; ></span>\frac{\mathrm{<span\; class="notranslate">\; z</span>}}{\mathrm{<span\; class="notranslate">\; h</span>}}<span\; class="notranslate">\; (</span>\mathrm{<span\; class="notranslate">\; 1</span>}<span\; class="notranslate">\; -</span>\frac{\mathrm{<span\; class="notranslate">\; z</span>}}{\mathrm{<span\; class="notranslate">\; h</span>}}<span\; class="notranslate">\; )</span><span\; class="notranslate">\; ,</span>$

Where <V _m > is the average velocity of the carrier medium, H is the height of the reaction tank, and V _m is the velocity at which the distance between the position of the carrier medium and the bottom of the reaction tank is z. It should be pointed out that the above parabolic velocity distribution is only an approximation of the velocity distribution under the condition that the length and width of the reaction pool are much larger than the height of the reaction pool. We therefore use the term approximately parabolic distribution in this patent text. Along the width of the bath, the parabolic distribution of the above formula is more accurate for positions in the middle of the width of the bath than for positions at the ends. Similarly, along the length of the bath, the parabolic distribution of the above formula is more accurate for positions in the middle of the length of the bath than for positions at the ends. The average speed can be calculated according to the following formula:

Average velocity <V _m >=(flow velocity)/(reaction cell width×reaction cell height or thickness)

Thus, structural features of the reaction cell that affect the velocity distribution of fluid flow include: reaction cell width, reaction cell height (or reaction cell thickness), and reaction cell length. Reactor cells of different sizes and geometries will result in different velocity profiles as the fluid passes through the cell. Parameters that determine the velocity distribution of fluid flow include, but are not limited to, reaction cell geometry; compression and expansion of fluid flow paths, for example, those opposing reaction cell walls in a non-parallel position, or proper placement of obstacles or wings ; the surface roughness of the reaction cell wall; the structural features of the reaction cell wall cause periodic or non-periodic modification of the fluid thickness, including electrode units and other surface structural configurations, and the geometry of the reaction cell can be, for example, rectangular, circular, wedge etc.

In the above method, the separated species can be discharged from one of the plurality of outlet ports of the reaction cell according to the position in the velocity distribution. In an example of a preferred entity, the gravitational force acting on the substance acts in a direction perpendicular to the flow of the carrier medium in the reaction cell.

The acoustic field-field flow separation, acoustic field-electrophoretic-field flow separation and acoustic field-dielectric-field flow separation methods described herein can be used to identify any substance. In a specific example of entities, the substances to be identified are cells, organelles, viruses, molecules or their aggregates or complexes. Recognizable cells include, but are not limited to, animal cells, plant cells, fungal cells, bacteria, cultured cells, or recombinant cells. Identifiable organelles include, but are not limited to, the nucleus, mitochondria, chloroplasts, ribosomes, rough endoplasmic reticulum, Golgi apparatus, lysosomes, proteasomes, secretory vesicles, vacuoles, or microsomes. Recognizable molecules include inorganic such as ions, organic molecules or complexes thereof. Recognized ions include, but are not limited to, sodium, potassium, magnesium, calcium, aluminum, chlorine, iron, copper, zinc, manganese, cobalt, iodine, molybdenum, vanadium, nickel, chromium, fluorine, silicon, tin, boron, arsenic ion. Recognizable organic molecules include, but are not limited to, amino acids, peptides, proteins, nucleosides, nucleotides, oligonucleotides, nucleic acids, vitamins, monosaccharides, oligosaccharides, carbohydrates, lipids, or complexes thereof. The substances to be identified can be of any size. Preferably, however, the size of the substance to be identified is between 0.01 microns and 1000 microns.

Description of drawings

Fig. 1 is a schematic diagram of a sound field-field flow separation reaction cell with a rectangular channel cut in the middle. At the same time, the working principle of sound field-field flow separation is illustrated.

Fig. 2 is a schematic diagram of an acoustic field-field flow separation reaction cell with an elliptical channel cut in the middle.

Fig. 3 is a schematic diagram of an elliptical sound field-field flow separation reaction cell with a channel cut in the middle.

Fig. 4 is a schematic diagram of an acoustic field-field flow separation reaction cell with multiple outlets at the outlet end of the reaction cell.

Fig. 5 is a schematic diagram of an acoustic field-electrophoresis-field flow separation reaction cell with a rectangular channel cut in the middle. At the same time, the working principle of acoustic field-electrophoresis-field flow separation is illustrated.

Fig. 6 is an electrode array that can be used in the acoustic field-electrophoresis-field flow separation device. (A) Mutually dislocated electrode array. (B) Mutually dislocated castle electrode array.

Fig. 7 is a schematic diagram of the acoustic field-dielectric-field flow separation reaction cell with a rectangular channel cut in the middle, and also illustrates the working principle of the acoustic field-dielectric-field flow separation.

Figure 8 is an electrode array that can be used in the acoustic field-dielectric-field current separation device. (A) Mutually displaced electrode array with triangular-shaped ends. (B) Mutually displaced electrode arrays with periodic arc-shaped ends.

Fig. 9 schematically illustrates the "batch mode working principle" using acoustic field-field flow separation, acoustic field-electrophoresis-field flow separation and acoustic field-dielectric-field flow separation devices. (A) As a result of the applied force during the "relaxation" process, different types of particles are distributed at different equilibrium heights. (B) Different types of particles distributed at different equilibrium heights move along the reaction cell at different speeds under the influence of fluid flow in the reaction cell.

Figure 10 schematically illustrates the "batch mode working principle" using acoustic field-field flow separation, acoustic field-electrophoresis-field flow separation and acoustic field-dielectric-field flow separation devices. The particles in the carrier medium are continuously injected into the reaction tank, and are continuously distributed at different heights in the carrier medium, and then discharged from different outlet ports.

Modes for Carrying Out the Invention

A. Definition

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by anyone with ordinary skill in the field of the invention. All patents, applications, published applications, and other publications and sequences in GenBank and databases cited in any part of this application are incorporated by reference into this patent.

Herein, "substance" refers to particulate matter, soluble matter or combinations thereof.

Here, "electrode unit (or electrode)" refers to a structure made of high-conductivity material to which a constant or approximately constant voltage can be applied. Approximately constant means that the value of the voltage drop across such a conductivity structure is so small that an electrical signal applied to an elliptical, triangular, rectangular, hexagonal or other geometrically shaped structure can generate a sufficiently strong electric field. Typical high-conductivity materials such as metal films (such as gold, platinum, titanium, chromium, etc.), semiconductor materials such as silicon-doped (such as silicon doped with phosphorus, arsenic, antimony, aluminum, gallium or indium), and other high-conductivity materials Material. For the purposes of the present invention, a material with high electrical conductivity refers to an electrical conductivity substantially greater than that of the carrier medium used for separation (eg, the conductive material has twice or more the conductivity of the carrier medium). The terms described in the present specification and claims must be understood to include all electrode configurations.

The "electrode array" mentioned here refers to a collection of multiple electrodes. In the electrode array, the relative positions of individual electrodes can be arranged according to a certain specified geometric relationship. For example, mutually dislocated electrode arrays, mutually dislocated castle electrode arrays, polynomial electrode arrays, mutually dislocated electrode arrays with periodic triangular tops (or arc tops, rectangular tops) or similar electrode arrays. An electrode array may comprise a plurality of electrodes of the same or different geometries.

The term "perpendicular to the conveying direction" of the carrier medium flowing through the reaction cell herein refers to a direction that is substantially not opposite to or parallel to the flow direction. For example, when a carrier medium flows along the length of a rectangular reaction cell, the direction "perpendicular to the direction of transport" may be the direction across the width of the reaction cell, the direction across the height of the reaction cell, or a direction that lies between In a certain plane parallel to the plane determined by the width and height of the reaction pool, or in any direction, as long as it is not opposite to the flow direction of the carrier medium. Usually, the included angle between the transport direction and the "perpendicular to the transport direction" is between 45° and 135°, preferably the included angle is between 80° and 100°, more preferably between 85° and 95°.

A "separation collector (or collection pool, collection means)" includes storage and collection means for holding separated or differentiated or identified substances.

"Identification means" means those which can be used to identify those isolated or identified or distinguished substances. The identification device may be a particle counter for recording the number of particles or the time the particles have left the reaction cell. The identification device can also be an assay device for more in-depth analysis and separation.

"Piezoelectric sensor" refers to a piezoelectric material member that generates an electric field when it changes in dimension due to an imposed mechanical force. Mechanical pressure can be obtained by applying an electrical signal. In this invention, we apply an alternating current signal to the piezoelectric sensor to generate alternating mechanical pressure, which is coupled into an acoustic wave into the carrier medium for separation and identification of substances.

"Structural features of the reaction pool" refers to the structural features of the reaction pool, including but not limited to the geometric shape, size, structure and position of the reaction pools (such as top cover, bottom surface, side wall) of the reaction pool.

"The carrier medium moves at different speeds at different positions in the reaction tank" means that the structural features of the reaction tank are designed or selected so that the carrier medium moves at different speeds at at least two positions in the reaction tank. It is generally not necessary for the carrier medium to move at different speeds at all locations in the reaction cell. In many cases, as long as the speed of the carrier medium is the same at a certain height (or width) or on a certain plane perpendicular to the direction of motion, it is the same as that of other heights (or widths) or other parallel planes. The speed of movement is different as it can be.

"Matter is transported to a position in a particular direction" means that matter is moved to a precise position in a direction of interest due to the force exerted on it. The location here is determined as a location or point in that direction. For example, in a rectangular reaction cell, the reaction cell consists of a top cover and a bottom surface and a thin gasket separating them. A shallow rectangular channel is carved in the middle of the thin gasket. Move in the length direction. Electrodes and piezoelectric sensors are mounted on the top and/or bottom of the reaction cell. When an electrical signal is applied to the electrodes and the piezoelectric sensor, an acoustic field force and a dielectric force will be generated to act on the substance in the carrier medium. These forces have components along the vertical direction, normal to the direction of motion. These components cause matter to move to different positions along the vertical direction. For example, when an electrical signal is applied to excite the electrodes and the piezoelectric sensor, the substance that was originally located close to the bottom of the reaction cell will move to a certain height from the bottom. The force used to change the position of matter in the present invention includes acoustic field force, electrophoretic force, dielectric force, gravity, fluid lift force, thermal diffusion force. For a "substance" whose position is affected by thermal diffusivity, the position of the "substance" refers to a distribution or a concentration distribution along a certain direction. For substances whose position is not affected by thermal diffusivity, the position of "substance" in a certain direction refers to the specific position of "substance" in this direction.

"Displacement within a velocity profile" refers to mass displacement within the velocity profile of a carrier medium moving in a reaction cell. The displacement here refers to the displacement along the direction of the velocity distribution curve. Matter in the fast-moving part of the velocity distribution will move faster than material in the slow-moving part of the velocity distribution. For substances whose positions are affected by thermal diffusion forces, the "substance" displacement within the velocity distribution refers to the concentration distribution of the species within the velocity distribution's reference frame. For "matter" whose position is not affected by the thermal diffusion force, the material displacement within the velocity distribution refers to the material displacement within the reference frame of the velocity distribution.

"An equilibrium position where a substance is displaced in a certain direction" refers to an equilibrium position of interest in which a substance is displaced in a certain direction under the influence of a force acting thereon. The equilibrium position refers to a place or point in this direction. A position of equilibrium is one where the net force on the matter is zero or nearly zero, such that the matter remains in that position. For a substance whose position is affected by thermal diffusion, the equilibrium position of a "substance" in a certain direction refers to the distribution of equilibrium positions of the substance in that direction. For substances whose position is not affected by thermal diffusion, the equilibrium position of the "matter" in a certain direction refers to the position of the "matter" in this direction when the material is balanced by force.

"The length of the reaction well is sufficiently large relative to the width and height" means that, irrespective of the actual shape of the reaction well, the typical length of the reaction well is at least twice the typical width and height of the reaction well. Preferably the typical length is at least three times the typical width and height, more suitably the typical length of the reaction well is at least five times the typical width and height. While the width and height of the reaction well may be the same or approximately the same, preferably one is longer than the other, such as the width being substantially greater than the height, for example, a typical width of a reaction well may be at least two, three times, or Five times bigger.

For the clarity of the description, without being limited by the manner, the detailed description of the invention is divided into the following two parts.

B. Exemplary installation

B.1. Acoustic field-field flow separation reaction cell

Figure 1 shows a physical example of an acoustic field-field-flow separation reaction tank and the operating principle of the acoustic field-field-flow separation. The reaction cell has a top cover 10 and a bottom surface 20 . The top and bottom surfaces are separated by a gasket or partition 30 with a rectangular channel 40 engraved therein. The channel 40 ends in a tapered shape. For clarity, the top cover 10, gasket 30 and bottom surface 20 are shown separately. When in use, these components are connected together to form an acoustic field-field flow separation reaction pool. The inlet 50 and the outlet 60 are respectively located on the top cover and the bottom surface, the inlet port and the outlet port of the reaction pool. The inlet 50 is connected to an injection device 70, which can introduce the carrier medium and the substances to be separated into the reaction cell. The injection device can be a syringe pump equipped with an injection valve (Wang, et al., 1998). The outlet 60 is connected with a collecting or testing device 80 and can be used for testing the separated and differentiated substances after the acoustic field-field flow separation. The collection or assay device may be a particle counter, a flow cytometer, or a separation collector.

In the drawing of the solid example, the entire bottom surface 20 is a piezoelectric sensor. The upper surface 90 and the lower surface 100 of the bottom surface 20 are coated with metal film or other conductive materials. The AC signal output from the signal generator 110 is applied to the upper surface 90 and the lower surface 100 of the piezoelectric sensor 20 to generate sound waves in directions perpendicular to the surfaces 90 and 100 . The sound waves emitted by the piezoelectric sensor are reflected onto the top cover 10 . The sound waves emitted by the piezoelectric sensor and the sound waves reflected by the top cover are superimposed to form a synthetic sound field in the reaction cell. A synthetic sound field can have two components, a standing wave component and a traveling wave component. The ratio of the standing wave component and the traveling wave component depends on the height of the reaction cell (the distance between the top cover and the bottom surface), the wavelength of the sound wave, the acoustic properties of the top cover 10 and the bottom surface 20, and the attenuation coefficient of the sound wave in the carrier medium. A solid example is that the height of the reaction cell is half the wavelength of the standing wave in which a standing wave will form. There is a sound pressure node in the center plane of the reaction cell. Another example is where the height of the reaction cell is greater than or less than half the wavelength of the standing wave.

In the example of FIG. 1 , the bottom surface 20 corresponds to a piezoelectric sensor. Variations are possible for mounting one or more piezoelectric sensors along various portions of the reaction cell. Can be mounted on top or bottom. In the case of bottom-mounted piezoelectric sensors, the sensors can be bonded to the lower surface of a solid plate such that the plate forms the bottom of the reaction cell. Piezoelectric transducers generate sound waves that are coupled via a solid plate to the carrier medium in the reaction cell. Similarly for the case where the piezoelectric sensor is mounted on the top cover, the sensor can be bonded to the upper surface of a solid plate which can form the top cover of the reaction cell. Piezoelectric transducers generate sound waves that are coupled via a solid plate to the carrier medium in the reaction cell. The field flow separation reaction cell in Figure 1 includes a piezoelectric sensor. Multiple piezoelectric sensors can be used in one reaction cell. These sensors can be connected in series on the top cover or the bottom of the reaction cell, or both the top cover and the bottom, to form a piezoelectric sensor array. The same or different electrical signals can be applied to these piezoelectric transducers to generate sound waves in the reaction cell.

Substances introduced into the reaction cell are subject to different forces in the reaction cell. We consider a situation where the introduced substance is a fine particle and the reaction cell is placed flat. These forces are:

The sound field radiation force F _acoustic 120 in the vertical direction, pointing to or away from the top cover (bottom surface), depends on factors related to particle and suspended carrier density and acoustic impedance. The acoustic field radiation force F _acoustic 120 may only be one component of the overall acoustic field force acting on the particle.

The gravitational force F _G 130 that lifts or settles the particles depends on the relative densities of the particles and the suspended carrier. Fluid lift, F _Lift 140, tends to push particles out of the reaction cell. Although fluid lift has been studied differently theoretically and experimentally, its nature remains a problem (Williams et al., 1992; 1994; 1996; 1997). Regardless, the generally accepted view is that this force plays a large role when the particles are very close to the cell wall (eg, less than 5 um from the wall in a 200 um thick cell). Recent studies on dielectric-field current separation (Huang et al., 1997; Wang et al., 1998) show that this force has limited effect on dielectric-field current separation.

These three forces act on the particle to move it toward the equilibrium position, where the forces balance each other, and the net force acting on a single particle is zero in the vertical direction, which is:

F _acoustic (z)+F _Lift (z)-F _G ＝0

Particles with different properties (eg, size, geometry, density, acoustic impedance) have different equilibrium positions at different heights. For example, particles 150 and 160 are transferred to different heights from the bottom of the reaction tank. Velocity profile 170 occurs when a carrier medium is introduced through inlet 50 to create flow. The direction of motion in this example is parallel to the top and bottom surfaces, pointing from the inlet end of the reaction cell to the outlet end. The carrier medium has different velocities at different locations. For the example in Figure 1, when the length of the reaction cell (i.e. the length of the etched channel) and width (i.e. the width of the channel) are much greater than the height (i.e. the distance between the top cover and the bottom surface), the carrier farther away from the channel wall formed by the gasket The velocity of the medium obeys an approximate parabolic velocity distribution in the vertical direction,

${\mathrm{<span\; class="notranslate">\; V</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}<span\; class="notranslate">\; =</span>\mathrm{<span\; class="notranslate">\; 6</span>}<span\; class="notranslate">\; <</span>{\mathrm{<span\; class="notranslate">\; V</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}<span\; class="notranslate">\; ></span>\frac{\mathrm{<span\; class="notranslate">\; z</span>}}{\mathrm{<span\; class="notranslate">\; h</span>}}<span\; class="notranslate">\; [</span>\mathrm{<span\; class="notranslate">\; 1</span>}<span\; class="notranslate">\; -</span>\frac{\mathrm{<span\; class="notranslate">\; z</span>}}{\mathrm{<span\; class="notranslate">\; h</span>}}<span\; class="notranslate">\; ]</span>$

Here V _m is the velocity of the carrier at the height z from the bottom of the reaction tank, <V _m > is the average velocity of the carrier, and H is the height of the reaction tank. In this way, the carrier medium in the reaction pool forms an approximate parabolic velocity distribution in the vertical direction. The particles 150 and 160 can be differentiated according to their heights h1 and h2 in a direction perpendicular to the direction of movement of the carrier medium. In addition, the particles 150 and 160 can be further differentiated according to the vertical position on the velocity distribution. Still further, particles 150 and 160 pass through the reaction cell at different speeds. If the particles 150 and 160 are introduced into the reaction cell at approximately the same time at different velocities, the particles 150 and 160 will reside in the reaction cell for different times because of the different transport velocities. Particles of different properties (e.g., size, density, geometry, acoustic impedance) can be transferred to different vertical positions, can be differentiated according to the transferred position in the vertical direction or within the velocity distribution, can be based on the particle's passage through the reaction cell speed or dwell time. Particles of different properties can be divided into subpopulations. Additionally, if the outlets are distributed vertically at the end of the reaction cell, particles of different heights can be divided into subpopulations as they exit the reaction cell through different outlets.

In the above discussion, consideration is given to the case where the reaction cell is laid flat. However, the reaction cell can be positioned in any direction or at any angle to the horizontal. In these cases we still consider that the forces acting on the particles to be discriminated are mainly in a direction perpendicular to the direction of motion of the carrier medium. The difference between these cases and the horizontal placement of the reaction cells described above is gravity. In the above case, the angle between the acting direction of gravity and the moving direction of the carrier is 90 degrees. In the case that the reaction cell is not laid flat, the angle between the direction of gravity and the direction of movement of the carrier may not be 90 degrees. In this way, only one component of gravity needs to be considered when analyzing the force exerted on the substance to be distinguished along the direction with an angle of 90 degrees to the direction of motion of the carrier medium.

The velocity distribution of the reaction cell in Figure 1 is determined by the structural characteristics of the reaction cell. When the length and width of the reaction pool are much greater than the height, there is a parabolic or nearly parabolic velocity distribution along the vertical direction of the reaction pool. The reason for the approximate parabola is that the velocity distribution close to the gasket wall is not a "parabolic distribution". When the width of the reaction cell is close to the height, the velocity of the carrier medium will follow other velocity distributions than the "parabolic distribution" mentioned above. Furthermore, the top and bottom surfaces are considered flat and parallel to each other in the above discussion. The velocity profile of the carrier medium can differ from the "approximately parabolic distribution" described above when the top and/or bottom surfaces are uneven or not parallel to each other, or if the top or bottom surfaces are altered by structural elements of different thickness.

In order to generate different velocity profiles of the carrier medium, channels of different shapes can be etched into the gasket 30 between the top cover 10 and the bottom surface 20 . For example, the channel 40 in the intermediate flow separation reaction cell in Fig. 2 is elliptical. When the carrier medium passes through such a reaction cell, the velocity profile differs from that in FIG. 1 . Similarly, a unique velocity profile is created when the carrier medium passes through the channels in the reaction cell of Figure 3.

In the above discussion about the field flow separation reaction cell, we mainly focused on the distinction of particles and ignored the influence of thermal diffusion effect. For the fine substances to be separated in the field flow separation reaction cell, it is necessary to consider the thermal diffusion force. In this case, the action of the force is shifted in a certain direction or the position of the matter within the velocity distribution refers to the distribution of matter under the influence of the action of the force. The substance distribution along a certain direction or within the velocity distribution refers to the concentration distribution of the substance along a certain direction or within the velocity distribution.

The field flow separation reaction cell can have one or more inlets to introduce the substances to be separated and the carrier medium. The field flow separation reaction cell can have one or more outlets to lead out the separated substances and carrier media. The inlet and outlet can be located on the top cover and/or the bottom surface. The inlets and outlets can be holes (from a few micrometers to millimeters in diameter) drilled in the top and/or bottom surface. PEEK, plastic, or metal tubing can be inserted into the well to connect the reaction cell to external fluid pathways, such as injection or collection devices. The inlets and outlets can also be channels (from a few micrometers to millimeters in width) drilled through the end of the reaction cell. A multi-pipe system arranged in a strip can be connected to the channel. In the reaction cell of Figure 4, a single inlet - orifice - is located at the bottom of the reaction cell. The two output ports 180 and 190, located both on the top and bottom, are channels cut in the wall. Most of the ductwork in the form of a ribbon acts as an outlet for connecting channels. The two outlets located at the top and bottom correspond to the separation devices used in field flow separations (Springston et al, 1987; Lee et al, 1989; Levin and Giddings, 1991).

B.2. Acoustic field-electrophoresis-flow separation reaction cell

Figure 5 shows a specific realization and operation principle of acoustic field-electrophoresis-field flow separation. The reaction cell has a top cover 210 and a bottom surface 220 . The top and bottom surfaces are separated by a gasket or partition 230 with a rectangular channel 240 carved into it. Channel 240 has a tapered end. Top cover 210, bottom surface 220, gasket 230, and channel 240 are shown separately for clarity. When in use, these components are connected together to form an acoustic field-electrophoresis-field flow separation reaction pool. The inlet 250 and the outlet 260 are respectively located on the top cover and the bottom surface, the inlet end and the outlet end of the reaction pool. The inlet 250 is connected with an injection device 270 for introducing the carrier medium and the substances to be separated into the reaction cell. The injection device can be a syringe pump incorporating an injection valve (Wang et al., 1998, Huang et al., 1999, Yang et al., 1999). Outlet 260 is associated with a collection or assay for the separation and separation of substances after assay field flow separation. The collection or assay device can be a particle counter, flow cytometer or separation collector.

In the example in Figure 5, the entire bottom surface is a piezoelectric sensor. The top surface 290 and the bottom surface 300 of the bottom surface 220 are coated with a layer of metal film or other conductive materials. The AC signal output from the signal generator 310 is applied to the upper surface 290 and the lower surface 300 of the piezoelectric sensor 220 to generate sound waves in directions perpendicular to the surfaces 290 and 300 . The sound waves emitted by the piezoelectric transducer are reflected to the top cover 210 . The sound waves emitted by the piezoelectric sensor and the sound waves reflected by the top cover are superimposed to form a synthetic sound field in the reaction cell. A synthetic sound field can have two components, a standing wave component and a traveling wave component. The ratio of the standing wave component to the traveling wave component depends on the height of the reaction cell (the distance between the top cover and the bottom surface), the wavelength of the sound wave, the acoustic properties of the top cover 210 and the bottom surface 220, and the attenuation coefficient of the sound wave in the carrier medium. A solid example is a reaction cell whose height is half the wavelength of the standing wave in which a standing wave will form. There is a sound pressure node in the center plane of the reaction cell. Another example is where the height of the reaction cell is greater than or less than half the wavelength of the standing wave.

In the example of FIG. 5, the bottom surface 220 corresponds to a piezoelectric sensor. Variations are possible for mounting one or more piezoelectric sensors along various portions of the reaction cell. Can be mounted on top or bottom. In the case of bottom-mounted piezoelectric sensors, the sensors can be bonded to the lower surface of a solid plate such that the plate forms the bottom of the reaction cell. Piezoelectric transducers generate sound waves that are coupled via a solid plate to the carrier medium in the reaction cell. Similarly for the case where the piezoelectric sensor is mounted on the top cover, the sensor can be bonded to the upper surface of a solid plate which can form the top cover of the reaction cell. Piezoelectric transducers generate sound waves that are coupled via a solid plate to the carrier medium in the reaction cell. The field flow separation reaction cell in Figure 5 includes a piezoelectric sensor. Multiple piezoelectric sensors can be used in one reaction cell. These sensors can be connected in series on the top cover or the bottom of the reaction cell, or both the top cover and the bottom, to form a piezoelectric sensor array. The same or different electrical signals can be applied to these piezoelectric transducers to generate sound waves in the reaction cell.

In FIG. 5 , the electrodes for generating an electric field in the reaction cell correspond to the upper surface 290 of the bottom surface 220 and the lower surface 295 of the top cover 210 . The upper surface 290 of the bottom surface 220 and the lower surface 295 of the top cover 210 are coated with a layer of metal film or other conductive materials. In this way, the DC signal or the low frequency AC signal generated by the signal generator 315 is applied between the surfaces 290 and 295 to generate an electric field perpendicular to the top cover and the bottom surface. Substances introduced into this electric field are subjected to electrophoretic forces that depend on the field strength and the effective charge of the substance.

In the example of FIG. 5, the upper surface 290 of the bottom surface 220 serves as an electrode for generating an electric field, and is also used to excite the piezoelectric sensor. Typically, the electrodes used to generate the electric field are different from the electrodes used to excite the piezoelectric sensor. Furthermore, the electrodes for generating the electric field may only partially cover the lower surface of the top cover or the upper surface of the bottom surface. Different electrode arrays can be applied on these surfaces (ie the lower surface of the top cover or the upper surface of the bottom surface). FIG. 6 shows an interdigitated electrode array 400 and an interdigitated castellation electrode array 410 . The electrodes can be distributed longitudinally (as shown in FIG. 6A or FIG. 6B ) or horizontally (ie, the electrodes in FIG. 6A are rotated by 90 degrees) along the reaction cell. Individual electrodes in the electrode array are connected to one of two electrical conductor buses 405 and 408 . When the signal from the signal source is connected to the electric wire bus, the electrodes are excited to generate an electric field. In those cases where an electrode array is used to generate an electric field, different electrical signal connection modes may be used for generating the electric field and for actuating the piezoelectric sensor. For example, when an electrode is used to generate an electric field in the reaction cell, the lower surface of the top cover forms a conductive plane, the lower surface of the bottom surface is coated with a metal film, and the upper surface is covered with a cross-shaped electrode array to form a piezoelectric sensor. The electrical signal to generate an electric field in the reaction cell can be applied across the lower surface of the top cover and either or both of the electrical wire buses (405 and 408 in FIG. 6A ) in the intersecting array. The electrical signal to generate an electric field in the reaction cell can be applied through the lower surface of the bottom surface and any one or both of the electrical wire buses (405 and 408 in FIG. 6A ) in the cross array.

The substances introduced into the acoustic field-electrophoresis-field flow separation reaction cell will be affected by different forces in the reaction cell. We consider a situation where the introduced substance is a fine particle and the reaction cell is placed flat. Referring to Figure 5, these forces are:

The sound field radiation force F _acoustic 320 in the vertical direction, pointing to or away from the top cover (bottom surface), depends on factors related to particle and suspended carrier density and acoustic impedance. The acoustic field radiation force F _acoustic 320 may only be a component of the total acoustic field force acting on the particle.

Electrophoretic force F _E 325 acting perpendicularly on charged particles. It depends on whether the particle is positively charged or negatively charged, and the direction of the DC electric field. The force is directed towards or away from the bottom of the reaction cell. This electrophoretic force F _E 325 may only be a component of the total electrophoretic force acting on the particle.

The gravitational force F _G 130 that lifts or settles the particles depends on the relative densities of the particles and the suspended carrier.

Fluid lift, F _Lift 340, tends to push particles out of the reaction cell. Although fluid lift has been studied differently theoretically and experimentally, its nature remains a problem (Williams et al., 1992; 1994; 1996; 1997). Regardless, the generally accepted view is that this force plays a large role when the particles are very close to the cell wall (eg, less than 5 um from the wall in a 200 um thick cell). Recent studies on dielectric-field current separation (Huang et al., 1997; Wang et al., 1998) show that this force has limited effect on dielectric-field current separation.

These forces act on the particle to move it to the equilibrium position, where the forces balance each other, and the net force acting on a single particle is zero in the vertical direction, that is:

F _acoustic (z)+F _Lift (z)+F _E (z)-F _G ＝0

Particles of different properties (eg, size, geometry, density, acoustic impedance) have different equilibrium positions at different heights. For example, particles 350 and 360 are transferred to different heights from the bottom of the reaction tank. Velocity profile 270 occurs when a carrier medium is introduced through inlet 250 to create flow. The direction of motion in this example is parallel to the top and bottom surfaces, pointing from the inlet end of the reaction cell to the outlet end. The carrier medium has different velocities at different locations. For the example in Figure 5, when the length of the reaction cell (i.e., the length of the etched channel) and the width (i.e., the width of the channel) are much greater than the height (i.e., the distance between the top cover and the bottom surface), the carrier farther away from the channel wall formed by the gasket The velocity of the medium obeys an approximate parabolic velocity distribution in the vertical direction,

${\mathrm{<span\; class="notranslate">\; V</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}<span\; class="notranslate">\; =</span>\mathrm{<span\; class="notranslate">\; 6</span>}<span\; class="notranslate">\; <</span>{\mathrm{<span\; class="notranslate">\; V</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}<span\; class="notranslate">\; ></span>\frac{\mathrm{<span\; class="notranslate">\; z</span>}}{\mathrm{<span\; class="notranslate">\; h</span>}}<span\; class="notranslate">\; [</span>\mathrm{<span\; class="notranslate">\; 1</span>}<span\; class="notranslate">\; -</span>\frac{\mathrm{<span\; class="notranslate">\; z</span>}}{\mathrm{<span\; class="notranslate">\; h</span>}}<span\; class="notranslate">\; ]</span>$

Here V _m is the velocity of the carrier at the height z from the bottom of the reaction tank, <V _m > is the average velocity of the carrier, and H is the height of the reaction tank. In this way, the carrier medium in the reaction pool forms an approximately parabolic velocity distribution in the vertical direction. The particles 350 and 360 can be differentiated according to their heights h1 and h2 in a direction perpendicular to the direction of movement of the carrier medium. In addition, the particles 350 and 360 can be further differentiated according to the vertical position on the velocity distribution. Still further, particles 350 and 360 pass through the reaction cell at different speeds. If the particles 350 and 360 are introduced into the reaction cell at approximately the same time at different velocities, the particles 350 and 360 will reside in the reaction cell for different times because of the different transport velocities. Particles of different properties (e.g., size, density, geometry, acoustic impedance) can be transferred to different vertical positions, can be differentiated according to the transferred position in the vertical direction or within the velocity distribution, can be based on the particle's passage through the reaction cell speed or dwell time. Particles of different properties can be divided into subpopulations. Additionally, if the outlets are distributed vertically at the end of the reaction cell, particles of different heights can be divided into subpopulations as they exit the reaction cell through different outlets.

In the above discussion, the reaction cell is considered to be placed horizontally, but in fact the reaction cell can be placed in any direction, or at any angle to the horizontal plane. In these cases we also need to take into account the forces acting on the objects to be separated at an angle of 90° to the direction of motion of the carrier medium. This is compared to the horizontal placement of the reaction cell discussed above, the difference is the difference in the effect of gravity. In the case discussed above, the direction of gravity acts at an angle of 90 degrees to the direction of motion of the carrier medium. When the reaction cell is not placed horizontally, the angle between the direction of gravity and the direction of movement of the carrier medium may not be 90 degrees. In this way, only one component of gravity needs to be considered when analyzing the force exerted on the substance to be distinguished along the direction with an angle of 90 degrees to the direction of motion of the carrier medium.

The velocity distribution in the rectangular reaction cell shown in Figure 5 depends on the structural properties of this reaction cell. When the length and width of the reaction pool are much greater than the height of the reaction pool, a parabolic or nearly parabolic velocity distribution along the vertical direction will appear in the reaction pool. The reason for the approximate parabolic shape is that the velocity distribution near the gasket wall is not parabolic. When the width of the reaction cell is as large as its height, the velocity of the carrier medium in the reaction cell will be different from the above distribution. Furthermore, in the above discussion the top and bottom surfaces are considered to be flat and parallel to each other. When the top and/or bottom surfaces are not flat, or when the top and bottom surfaces are not parallel to each other, or when the top and bottom surfaces vary with structural elements of different thicknesses, the velocity profile of the carrier medium will differ from the approximate Parabolic velocity distribution.

To create different velocity profiles of the carrier medium, the gasket 230 between the top cover 210 and the bottom surface 220 can be cut down the middle to form channels of other shapes. For example, the channel in the acoustic field-dielectric-field flow separation reaction cell can be an ellipse, similar to the channel in the acoustic field-field flow separation reaction cell shown in FIG. 2 . When the carrier medium passes through such a reaction cell, the velocity profile of the carrier medium will be different from that shown in FIG. 5 . Similarly, the channel in the acoustic field-field flow separation reaction cell shown in FIG. 3 can be used as an acoustic field-dielectric-field flow separation reaction cell. Such a reaction cell will create a unique velocity profile for the carrier medium as it passes through the channel.

The above discussion about the acoustic field-dielectric-field flow separation reaction cell mainly focuses on particle separation, ignoring the influence of thermal diffusion effect. For small objects to be separated in the acoustic-dielectric-field flow separation cell, thermal diffusion effects should be taken into account. Thus, the position of matter arranged in one direction or within a velocity distribution under the action of an external force refers to the distribution of matter under the action of an external force. The distribution of objects along a certain direction or a certain speed refers to the concentration distribution or position distribution of objects distributed along this direction or speed.

The acoustic field-dielectric-field flow separation reaction cell can have one or more inlets to input the objects to be separated and the carrier medium. The reaction cell can also have one or more outlets for the output of separated objects and carrier medium. Inlets and outlets can be placed on the top or/and bottom of the reaction tank. The inlet and outlet can be small holes (diameter as small as several microns or as large as several millimeters) drilled in the top and/or bottom of the reaction tank. PEEK or plastic, or metal tubing is inserted into the holes as the fluidic connection between the reaction cell and the external fluid circuit (eg diffuser or collection device). On the other hand, the inlet and outlet can also be a channel (width from microns to millimeters) at the outlet end of the reaction cell. Composite pipes, arranged in strips, can be connected with the above-mentioned passages. The exemplary acoustic field-field flow separation reaction cell shown in FIG. 4 can be used as an example of an acoustic field-dielectric-field flow separation reaction cell. At this time, a single input port (hole) is located on the bottom surface of the reaction cell. The two outlets 180 and 190 are located on the top and bottom of the reaction tank and are thin channels cut in the wall. A number of tubes arranged in ribbons are used to connect the outlets of these channels. The two outlets located at the top and bottom correspond to the separation devices used in field flow separations (Springston et al, 1987; Lee et al, 1989; Levin and Giddings, 1991).

B3. Acoustic field-dielectric-field flow separation reaction cell

Figure 7 shows the structure of an acoustic-dielectric-field-flow separation reaction cell and the working principle of the acoustic-dielectric-field-flow separation reaction cell. The reaction cell has a top cover 510 and a bottom surface 520, which are separated by a gasket 530, and a rectangular channel 540 is engraved on the 530. The two ends of the 540 are tapered. There is a piezoelectric sensor 525 on the underside. For clarity, the top cover 510, spacer 530, bottom surface 520, and piezoelectric sensor 525 are shown separately. In use, these components are bonded together to form an acoustic dielectric-field flow separation reaction cell. The inlet 550 and the outlet 560 are respectively located on the top cover and the bottom surface, that is, at the inlet end and the outlet end of the reaction pool respectively. The inlet 550 is connected to an injection device 570 which can introduce the carrier medium and the objects to be separated. The injection device can be a syringe pump with an injection valve (Wang et al., 1998; Huang et al., 1999; Yang et al., 1999). Outlet 560 is connected to a collection device 580 capable of identifying objects that have been separated by the ADF separation cell. The collecting device can be a particle counter, a flow cytometer or a split collector.

In Example 7, the piezoelectric sensor 525 corresponds to the entire bottom area of the reaction cell, and its upper surface 590 and lower surface 600 are coated with metal film and other conductive materials. The AC signal generated by the signal generator is applied to the upper surface 590 and the lower surface 600 of the piezoelectric sensor 525 to excite the piezoelectric sensor to generate sound waves in the direction perpendicular to 590 and 600 in the reaction cell. The sound wave emitted by the piezoelectric sensor is coupled with the bottom surface 520 into the reaction cell, and is reflected back by the top cover 510 . The superposition of the propagating wave of the piezoelectric sensor and the reflected wave of the top cover constitutes the entire sound field in the reaction cell. The full acoustic wave field has two components, the standing wave component and the traveling wave component. The ratio of the size of the two components depends on the height of the reaction cell (i.e. the distance between the top cover and the bottom surface of the reaction cell), the wavelength of the sound wave, and the acoustic characteristics of the top cover 510, the bottom surface 520 and the piezoelectric sensor 525, as well as the carrier medium and the sound wave. decay factor. In one implementation, the height of the reaction cell between the top cover and the bottom surface is half the wavelength of the standing wave, thus forming a standing wave in the reaction cell. There is a sound pressure node in the middle plane of the reaction cell. In another implementation, the reaction cell height is greater (or less) than half the wavelength of the standing wave.

In FIG. 7 , a piezoelectric sensor 525 is bonded to the bottom surface 520 of the reaction cell. There are various ways of placing one or more piezoelectric sensors along various sections of the reaction cell. 525 can be placed on top cover and/or bottom surface. When the piezoelectric sensor is placed on the top cover, it can be bonded to a solid piece from the top cover so that the solid piece forms the top cover of the reaction cell. Acoustic waves can be generated by piezoelectric transducers and coupled through solid plates into the carrier medium in the reaction cell. In addition, piezoelectric sensors can also be used directly as reaction cell top covers. When the piezoelectric sensor is placed on the bottom of the reaction pool, the sensor can be directly used as the bottom of the reaction pool, and the microelectrode element or array can be directly fabricated on the upper surface of the piezoelectric sensor. The acoustic dielectric-field flow separation reaction cell shown in Fig. 7 includes a piezoelectric sensor in the reaction cell. Multiple piezoelectric sensors can be placed in the same reaction cell. They can be placed in series on the top cover or bottom or both to form a piezoelectric sensor array. Multiple piezoelectric transducers can be excited by the same or different electrical signals to generate acoustic waves in the reaction cell.

In the example of FIG. 7 , the electrode elements for generating an electric field in the reaction cell correspond to the electrode array 545 on the upper surface of the bottom surface 520 . The signal generator 615 can be applied to the electrode array 545 to generate an AC signal to form a non-uniform electric field in the reaction cell. An object subjected to such an uneven electric field will be subjected to a dielectrophoretic force, which is related to the dielectric constant of the object and its surrounding medium, and also related to the uneven distribution of the electric field.

In the example of FIG. 7, an electrode array 545 is disposed on the upper surface of the bottom surface 520 to generate a non-uniform electric field. Generally, the electrode elements for generating the electric field may partially cover the lower surface of the top cover and the upper surface of the bottom surface or both. Electrode arrays of different configurations can be used on these surfaces (ie, the lower surface of the top cover and the upper surface of the bottom surface). The staggered electrode array 400 and the staggered castle electrode array 410 shown in FIGS. 6A and 6B can be used. The electrodes are distributed longitudinally (Fig. 6A, 6B, array 545 in Fig. 7) or transversely (Fig. 6A, 6B, electrode elements of array 545 in Fig. 7 are rotated 90 degrees) along the reaction cell. Similarly, as shown in Figure 8, an interlaced electrode array with periodic triangular (700) or arc-shaped electrodes (710) can also be used, and the electrodes can be vertically (Fig. 8A, 8B) or laterally (Fig. rotated 90 degrees) distribution. Individual electrode elements in the electrode arrays of Figures 8A, 8B are connected to one of two common electrical conductor buses 705,708. When the electrical signal sent by the signal source is applied to the electrical wire bus, the electrode elements are excited to generate an electric field.

The structure of the example shown in FIG. 7 is that the electrode array 545 is arranged on the bottom surface 520 of the reaction cell, and the piezoelectric sensor 525 is bonded on the bottom surface 520 . In other implementations, electrode arrays with different combined shapes can also be fabricated directly on the piezoelectric sensor. For example, PZT is a piezoelectric material that can be used in piezoelectric sensors. When its surface is ground to be smooth enough, micro-machining methods can be used to make micro-electrodes on its piezoelectric substrate. In the operation of constructing the acoustic dielectric-field flow separation reaction cell, the piezoelectric sensor with the microelectrode on the upper surface and the flat plate electrode on the lower surface can be used as the bottom surface of the acoustic dielectric-field flow separation reaction cell. Dielectrophoretic forces can be generated by applying electrical signals to the microelectrode array from the upper surface. Simultaneously, electrical signals can be applied to the upper electrode array (via electrical wire buses 705 and 708) and the lower electrode array to generate sound fields and forces. The advantage of this approach is that the electrode array used to generate the dielectrophoretic force is integrated on the surface of the piezoelectric sensor. This method of integrating the electrode array with the piezoelectric sensor is similar to the method of the acoustic field flow separation reaction cell shown in Fig. 5. The electrodes used to generate the electrophoretic force in Fig. 5 are on the upper surface of the piezoelectric sensor.

Objects introduced into the acoustic field-dielectric-field flow separation reaction cell will be subjected to various forces in the reaction cell. We consider the situation that the introduced objects are tiny particles and the reaction pool is placed horizontally. Referring to Figure 7, these forces are

Sound field radiation force F _acoustic 620: in the longitudinal direction or away from the top cover (bottom surface), it depends on a coefficient related to the density and acoustic impedance of particles and suspended media. The acoustic field radiation force F _acoustic 620 may only be a component of the total acoustic field force acting on the particles.

Dielectrophoretic force F _DEP 625: acts on polarized particles in the longitudinal direction. This force is directed towards or away from the electrode element, depending on whether the particle is more or less polarizable than the surrounding medium. This dielectrophoretic force F _DEP 625 may only be one component of the total dielectrophoretic force acting on the particle.

Gravity F _G 630: The force that suspends or deposits particles, related to the relative density of the particles and the surrounding medium.

Fluid Lift F _Lift 640: Attempts to drive particles away from the walls of the reaction tank. There have been various theoretical and experimental studies on fluid lift, but its nature remains unknown. The currently generally accepted view is that this force is only important when the particles are very close to the cell walls (eg, less than 5 microns distance in cells up to 200 microns thick). Some recent work on dielectric-field current separation (Huang et al., 1997; Wang et al., 1998) showed that this force plays a small role in the manipulation of dielectric-field current separation.

The above forces act on the particle, driving the particle to move towards the position where the total force is balanced, so the net force acting on a single particle in the longitudinal direction is 0, that is

F _acoustic (z)+F _Lift (z)+F _DEP (z)-F _G ＝0

Particles with different properties (such as size, geometry, density, dielectric constant, acoustic impedance) reach equilibrium at different heights. For example particles 650 and 660 are equilibrated at different heights (h1, h2) from the bottom of the reaction cell. When the carrier medium is injected into the reaction cell from the inlet 550 to form a fluid, a fluid velocity profile 670 is created. At this time, the direction of movement is parallel to the top cover and bottom surface of the reaction pool, and is directed from the inlet end to the outlet end. The carrier medium has different velocities at different locations in the reaction cell. As in the example in 7, when the length of the reaction cell (that is, the length of the channel cut in the middle) and the width of the reaction cell (that is, the width of the channel) are much greater than the height of the reaction cell (that is, the distance between the top cover and the bottom surface), the carrier medium is far away from the reaction cell The velocity at the location of the wall follows an approximately parabolic velocity profile in the longitudinal direction.

${\mathrm{<span\; class="notranslate">\; V</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}<span\; class="notranslate">\; =</span>\mathrm{<span\; class="notranslate">\; 6</span>}<span\; class="notranslate">\; <</span>{\mathrm{<span\; class="notranslate">\; V</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}<span\; class="notranslate">\; ></span>\frac{\mathrm{<span\; class="notranslate">\; z</span>}}{\mathrm{<span\; class="notranslate">\; h</span>}}<span\; class="notranslate">\; (</span>\mathrm{<span\; class="notranslate">\; 1</span>}<span\; class="notranslate">\; -</span>\frac{\mathrm{<span\; class="notranslate">\; z</span>}}{\mathrm{<span\; class="notranslate">\; h</span>}}<span\; class="notranslate">\; )</span>$

Where V _m is the velocity of the medium at the height Z from the bottom of the reaction tank, <V _m > is the average velocity of the medium, and H is the height of the reaction tank. This creates an approximately parabolic velocity profile in the longitudinal direction of the carrier medium in the reaction cell. The particles 550, 560 can thus be separated according to their different positions in the velocity distribution. Furthermore, particles 550, 560 pass through the reaction cell at different speeds. If the particles 550, 560 pass through the reaction cell at the same time, they will all leave the reaction cell at the same time due to their different moving speeds in the reaction cell. Particles with different properties (such as size, geometry, density, dielectric constant, acoustic impedance) can be placed at different positions in the longitudinal direction, can be separated according to the different positions in the longitudinal direction, and can be separated according to the reaction of the particles. The difference in cell speed, or the separation according to the difference in the time for particles to leave the reaction cell.

In the above discussion, the reaction cell is considered to be placed horizontally, but in fact the reaction cell can be placed in any direction, or at any angle to the horizontal plane. In these cases we also need to take into account the forces acting on the objects to be separated at an angle of 90° to the direction of motion of the carrier medium. This is compared to the horizontal placement of the reaction cell discussed above, the difference is the difference in the effect of gravity. In the case discussed above, the direction of gravity acts at an angle of 90 degrees to the direction of motion of the carrier medium. When the reaction cell is not placed horizontally, the angle between the direction of gravity and the direction of movement of the carrier medium may not be 90 degrees. In this way, only one component of gravity needs to be considered when analyzing the force exerted on the substance to be distinguished along the direction with an angle of 90 degrees to the direction of motion of the carrier medium.

The velocity distribution in the rectangular reaction cell shown in Figure 7 depends on the structural properties of this reaction cell. When the length and width of the reaction pool are much greater than the height of the reaction pool, a parabolic or nearly parabolic velocity distribution along the vertical direction will appear in the reaction pool. The reason for the approximate parabolic shape is that the velocity distribution near the gasket wall is not parabolic. When the width of the reaction cell is as large as its height, the velocity of the carrier medium in the reaction cell will be different from the above distribution. Furthermore, in the above discussion the top and bottom surfaces are considered to be flat and parallel to each other. When the top and/or bottom surfaces are not flat, or when the top and bottom surfaces are not parallel to each other, or when the top and bottom surfaces vary with structural elements of different thicknesses, the velocity profile of the carrier medium will differ from the approximate Parabolic velocity distribution.

To create different velocity profiles of the carrier medium, the gasket 530 between the top cover 510 and the bottom surface 520 can be cut down the middle to form channels of other shapes. For example, the channel in the acoustic field-dielectric-field flow separation reaction cell can be an ellipse, similar to the channel in the acoustic field-field flow separation reaction cell shown in FIG. 2 . When the carrier medium passes through such a reaction cell, the velocity profile of the carrier medium will be different from that shown in FIG. 7 . Similarly, the channel in the acoustic field-field flow separation reaction cell shown in FIG. 3 can be used as an acoustic field-dielectric-field flow separation reaction cell. Such a reaction cell will create a unique velocity profile for the carrier medium as it passes through the channel.

The above discussion about the acoustic field-dielectric-field flow separation reaction cell mainly focuses on particle separation, ignoring the influence of thermal diffusion effect. For small objects to be separated in the acoustic-dielectric-field flow separation cell, thermal diffusion effects should be taken into account. Thus, the position of matter arranged in one direction or within a velocity distribution under the action of an external force refers to the distribution of matter under the action of an external force. The distribution of objects along a certain direction or a certain speed refers to the concentration distribution or position distribution of objects distributed along this direction or speed.

The acoustic field-dielectric-field flow separation reaction cell can have one or more inlets to input the objects to be separated and the carrier medium. The reaction cell can also have one or more outlets for the output of separated objects and carrier medium. Inlets and outlets can be placed on the top or/and bottom of the reaction tank. The inlet and outlet can be small holes (diameter as small as several microns or as large as several millimeters) drilled in the top and/or bottom of the reaction tank. PEEK or plastic, or metal tubing is inserted into the holes as the fluidic connection between the reaction cell and the external fluid circuit (eg diffuser or collection device). On the other hand, the inlet and outlet can also be a channel (width from microns to millimeters) at the outlet end of the reaction cell. A plurality of tubes arranged in a strip shape are connected to the above-mentioned channels. The exemplary acoustic field-field flow separation reaction cell shown in FIG. 4 can be used as an example of an acoustic field-dielectric-field flow separation reaction cell. At this time, a single input port (hole) is located on the bottom surface of the reaction cell. The two outlets 180 and 190 are located on the top and bottom of the reaction tank and are thin channels cut in the wall. A number of tubes arranged in ribbons are used to connect the outlets of these channels. The two outlets located at the top and bottom correspond to the separation devices used in field flow separations (Springston et al, 1987; Lee et al, 1989; Levin and Giddings, 1991).

C, example method

C.1. Sound field-field flow separation, sound field-electrophoresis-field flow separation and sound field-dielectric-field flow separation batch operation

In batch operation, an acoustic-field-flow separation (or acoustic-electrophoretic-field-flow separation or acoustic-dielectric-field-flow separation) reaction cell is preloaded with carrier medium through the inlet. The mixed particle sample (eg, 100 uL) is injected or inserted into the medium in the reaction cell through the input port. For acoustic field-field flow separation, applying the appropriate acoustic field conditions, the particles will reach their equilibrium positions within a specified time under the influence of acoustic and other forces. The acoustic field in the reaction cell is obtained through appropriate electrical signals and piezoelectric transducers. For acoustic-electric-field flow separation, with appropriate electric and acoustic fields, the particles will reach their equilibrium positions within a specified time under the influence of acoustic, electrophoretic and other forces. For acoustic field-dielectric-field flow separation, with appropriate dielectrophoretic field and acoustic field, the particles reach their equilibrium positions within a period of time under the influence of acoustic field force, dielectrophoretic force and other forces. This step is called "relaxation" in typical field-flow separation processes (Giddings, 1981, Giddings, 1993). In the relaxation stage (FIG. 9A), particles with different properties are in different equilibrium positions in the reaction cell under the influence of applied force. For small-sized particles (or objects) where the thermal diffusion effect has an important influence on the equilibrium position of the particle, the equilibrium positions of different types of particles correspond to the equilibrium concentration distributions of different particle types. After the relaxation step, fluid flow is established and particles at different heights are driven through the reaction cell at different velocities. During this process, the conditions of the external field were kept constant. The particles with different properties were separated into multiple subgroups according to the time when the particles left the reaction cell ( FIG. 9B ). 9A, 9B show the batch process in a rectangular acoustic field-field flow separation (or electrophoretic-field flow separation, or dielectric-field flow separation) cross section. Figure 9A shows that during relaxation, particles 800 and 850 have been placed at different heights by the applied field. FIG. 9B shows that following the relaxation process, after establishing a fluid profile (ie velocity profile 780 ) in the reaction cell, particles 800 outnumber particles 850 and therefore leave the reaction cell at outlet 795 earlier. Fluid flow is achieved by injecting carrier medium into reaction cell inlet 790 .

Batch mode is a common mode of operation for most field flow separations. Particles may exit the reaction cell from a single outlet or from multiple outlets (eg, one at the top of the reaction cell and another at the bottom of the reaction cell). Two outlets on the top and bottom face can be used directly to collect the two separated parts. In some applications the relaxation step is unnecessary. Depending on the type of particle mixture pair, it is possible that the particles reach their equilibrium height very shortly after being introduced into the reaction cell. In this case, particles can be separated in batch mode without a relaxation step.

In this way, using acoustic field force in field flow separation to separate an object in a batch manner includes the following steps: 1) setting up the device described in the present invention; 2) introducing the carrier medium into the reaction cell of the device through the inlet port of the device until the carrier medium fills the entire reaction cell; 3) inject the sample containing the substance to be identified into the carrier medium in the reaction cell; 4) apply at least one electrical signal generated by the electrical signal generator to the piezoelectric sensor, wherein the excitation voltage The electrical sensor generates sound waves, thereby causing at least one acoustic field force to be exerted on the substance; 5) introducing the carrier medium into the reaction cell of the device through the inlet port of the device, wherein said introduction causes the carrier medium to pass through the reaction cell according to a velocity distribution; The substances in the carrier medium are distributed in the direction perpendicular to the carrier medium moving through the reaction pool, and the identification of the substances is realized according to the difference in the distribution position of the substances in the direction perpendicular to the carrier medium moving through the reaction pool. This acoustic field flow separation batch processing mode can be applied to various acoustic field-field flow separation devices described in the present invention.

In the above-mentioned acoustic field-field flow separation pair batch method, preferably before the carrier medium is introduced into the reaction cell to make it travel according to a certain velocity distribution, the piezoelectric sensor that can cause the acoustic field force on the object is first applied. The electric signal is used to place the object to be separated to the equilibrium position along the direction perpendicular to the direction of travel of the carrier medium in the reaction cell.

The application of electrophoretic force and acoustic field force in field flow separation to separate objects includes the following steps: 1) Establish the device described in the present invention. 2) introducing the carrier medium into the reaction cell of the device through the inlet port of the device until the carrier medium fills the entire reaction cell; 3) injecting the sample containing the substance to be identified into the carrier medium in the reaction cell; 4) applying the At least one electrical signal generated by an electrical signal generator, wherein the excitation electrode generates an electric field, thereby causing at least one electrophoretic force to be applied to the substance; 5) introducing the carrier medium into the device through the inlet port of the device In the reaction pool, wherein the introduction makes the carrier medium pass through the reaction pool according to the speed distribution; the substance in the carrier medium is distributed in the direction perpendicular to the direction of the carrier medium moving through the reaction pool, and the identification of the substance is based on the substance This is achieved by distributing positional differences in the direction perpendicular to the movement of the carrier medium through the reaction cell. Various sound field-electrophoresis-field flow separation devices described in the present invention can be applied in the batch processing mode of sound field-electrophoresis-field flow separation.

In the above-mentioned acoustic field-electricity-field flow separation pair batch process, preferably before the carrier medium is introduced into the reaction cell to make it travel according to a certain velocity distribution, the electrodes capable of inducing dielectrophoretic forces on said objects An electric signal is applied to the element and the piezoelectric sensor causing the acoustic field force to place the object to be separated to an equilibrium position in a direction perpendicular to the direction of travel of the carrier medium in the reaction cell.

The application of dielectrophoretic and acoustic field forces in field flow separation to separate objects in a batch manner involves the following steps: A) building the device described in the present invention; B) the reaction of introducing the carrier medium into the device through the inlet port of the device until the carrier medium fills the entire reaction cell; C) inject the sample containing the substance to be identified into the carrier medium in the reaction cell; D) apply at least one electrical signal generated by the electrical signal generator to the electrode, wherein the excitation electrode generating an electric field, thereby causing at least one dielectrophoretic force to be applied to said substance; E) applying at least another electrical signal generated by an electrical signal generator to the piezoelectric sensor, wherein said excited piezoelectric sensor generating sound waves, thereby causing at least one acoustic field force to be exerted on said substance; F) introducing the carrier medium into the reaction cell of the device through the inlet port of the device, wherein said introduction causes the carrier medium to pass through the reaction cell according to the velocity distribution The substances in the carrier medium are distributed in the direction perpendicular to the carrier medium moving through the reaction pool, and the identification of the substance is realized according to the difference in the distribution position of the substance in the direction perpendicular to the carrier medium moving through the reaction pool . Any device using the above invention can perform a batch operation of acoustic field-dielectric field flow separation.

In the above-mentioned "batch processing" operation using acoustic field-dielectric-field flow separation, it is best to first generate a dielectric force by applying an electrical signal to the electrode to be applied to the substance to be identified, and to generate an acoustic field by applying an electrical signal to the piezoelectric sensor A force is applied to the substance to be identified so that the substance to be identified is distributed in a direction perpendicular to the flow of the fluid and in equilibrium before being introduced into the medium to form a velocity distribution (step f).

In the above-mentioned batch operation using acoustic-electrophoretic-field-flow separation or acoustic-dielectric-field-flow separation, it is also preferable to apply different electrical signals to generate acoustic field force, electrophoretic force, or dielectrophoretic force .

In the above batch operation using acoustic field-electrophoresis-field-flow separation or acoustic-dielectric-field-flow separation, the acoustic field force, electrophoretic force or dielectrophoretic force can be generated simultaneously or sequentially.

C.2. Continuous mode operation of acoustic field-field-flow separation, acoustic field-electrophoresis-field-flow separation, and acoustic field-dielectric-field-flow separation

In continuous mode operation, the particulate mixed sample is continuously introduced into the reaction cell of acoustic field-field flow separation (or acoustic field-electrophoresis-field flow separation, acoustic field-dielectric-field flow separation) through the inlet port. For acoustic field-field flow separation, under the action of an appropriate acoustic field, the particles will be in an equilibrium position in the reaction cell under the action of the acoustic field force and other forces (such as gravity, fluid lift, etc.). The acoustic field force in the reaction cell is obtained by applying an appropriate electrical signal to the piezoelectric sensor. For acoustic field-electrophoresis-field flow separation, under the action of an appropriate electric field and acoustic field, the particles will be in an equilibrium position in the reaction cell under the action of the acoustic field force, electrophoretic force and other forces (such as gravity, fluid lift, etc.) . For the separation of acoustic field-dielectric-field flow, under the action of appropriate dielectric electric field and acoustic field, the particles will react under the action of acoustic field force, dielectrophoretic force and other forces (such as gravity, fluid lift, etc.). The pool is in equilibrium. Two kinds of particle samples are taken for separation and analysis in the acoustic field-field flow separation (or acoustic field-electrophoresis-field flow separation, acoustic field-dielectric-field flow separation) reaction cell. In the acoustic field-field flow separation (or acoustic field-electrophoresis-field flow separation, acoustic field-dielectric-field flow separation) reaction cell, the particles at a higher equilibrium position will flow out from the higher position of the outlet, and in the reaction cell at a higher position Particles in the low equilibrium position will flow out from the lower position of the outlet. In this way, the particle mixture is continuously divided into two layers. Clearly, there is a need to separate the fluids at the outlet of the reaction cell. (Springston et al, 1987; Lee et al, 1989; Levin and Giddings, 1991).

Multiple field flows can be collected if there are multiple outlets at the top and bottom of the tank. In the example of Figure 10, there is one outlet 870 at the top of the reaction cell and two outlets 880 and 890 at the bottom. In this way, this reaction cell can be used to separate three or more particles. The reaction cell can be an acoustic field-field flow separation reaction cell, or an acoustic field-electrophoresis-field flow separation, sound field-dielectric-field flow separation reaction cell. A fluid velocity profile 860 exists within the reaction cell.

Due to the various forces applied, the particles 900 are in the highest position during the flow through the reaction cell, and thus flow out from the top outlet 870 . However, due to various forces applied, the particles 910 are at the lowest position during the process of flowing through the reaction cell, and thus flow out from the outlet 880 at the bottom. Another particle 920 is in the middle so that it exits another outlet 890 at the bottom. This operation corresponds to a field flow separation system using a separation device. (Springston et al, 1987; Lee et al, 1989; Levin and Giddings, 1991).

In field flow separation, the continuous mode manipulation of separating substances by applying acoustic field force includes the following steps: a) obtaining an acoustic field-field flow separation device described in the present invention; B) passing through the inlet port of the device will contain The carrier medium of the substance to be identified is introduced into the reaction cell of the device, wherein the introduction makes the carrier medium pass through the reaction cell according to the speed distribution; C) the piezoelectric sensor is excited by applying at least one electrical signal generated by the electrical signal generator to generate Acoustic waves such that at least one acoustic field force exerted on a substance in said carrier medium has a component perpendicular to the direction of movement of the carrier medium through the reaction cell. The substances in the carrier medium are distributed in the direction perpendicular to the carrier medium moving through the reaction pool, and the substances are identified according to the difference in distribution position of the substances in the direction perpendicular to the carrier medium moving through the reaction pool. This continuous mode of sound field-field flow separation can be used in any sound field-field flow separation device.

The continuous mode manipulation of separating substances by applying acoustic field force in the acoustic field-electrophoresis-field flow separation includes the following steps: A) an acoustic field-electrophoresis-field flow separation device described in the present invention. B) introducing the carrier medium into the reaction cell of the device through the inlet port of the device, wherein the introduction causes the carrier medium to pass through the reaction cell according to the velocity distribution; C) by applying at least one path generated by an electrical signal generator to the electrodes The electric signal is excited to generate an electric field, thereby causing at least one electrophoretic force applied to the substance in the carrier medium to have a component perpendicular to the direction in which the carrier medium moves through the reaction cell; D) by applying an electric signal generator to the piezoelectric sensor The generated at least one other electrical signal excites an acoustic field such that at least one acoustic field force exerted on a substance in said carrier medium has a component perpendicular to the direction of movement of the carrier medium through the reaction cell. The substances in the carrier medium are distributed in the direction perpendicular to the carrier medium moving through the reaction pool, and the substances are identified according to the difference in distribution position of the substances in the direction perpendicular to the carrier medium moving through the reaction pool. This continuous mode of acoustic field-electrophoresis-field flow separation can be used in any acoustic field-electrophoresis-field flow separation device.

In the acoustic field-dielectrophoresis-field flow separation, the continuous mode manipulation of separating substances by applying the acoustic field force includes the following steps: A) Obtain an acoustic field-dielectric-field flow separation device described in the present invention. B) introducing the carrier medium into the reaction cell of the device through the inlet port of the device, wherein the introduction makes the carrier medium pass through the reaction cell according to the velocity distribution; C) applying at least one electric signal generated by the electric signal generator to the electrode signal, wherein the excitation electrode produces a non-uniform electric field, thereby causing at least one dielectrophoretic force exerted on the substance to have a component perpendicular to the direction of the carrier medium moving through the reaction cell; D) for pressure The electrical sensor applies at least another electrical signal generated by an electrical signal generator, wherein said excited piezoelectric sensor generates an acoustic field, thereby causing at least one acoustic field force applied to said substance to have a force perpendicular to the carrier medium The component moving through the direction of the reaction pool; the substance in the carrier medium is distributed in the direction perpendicular to the direction of the carrier medium moving through the reaction pool, according to the distribution position of the substance moving through the direction of the reaction pool perpendicular to the carrier medium Differently identify substances. This continuous mode of sound field-field flow separation can be used in any sound field-field flow separation device. This continuous mode of acoustic field-dielectrophoresis-field flow separation can be used in any acoustic field-dielectrophoresis-field flow separation device.

D. force

Although we do not wish to be bound by the theory and mechanisms described here, the following describe several current devices and methods utilizing the various forces.

D.1. Radiation force of sound field

Acoustic radiation force is a non-contact force that can be used in fluids to capture, manipulate, and move particles. The operations on particles using the acoustic field force generated by ultrasonic waves include collecting red blood cells (Yasuda et al, 1997), collecting micron-sized polyethylene particles (0.3 to 10 microns in diameter, Yasuda and Kamakura, 1997), and collecting DNA molecules (Yasuda et al. al, 1996C), large-scale, semi-continuous collection and sedimentation of cells (Pui, et al, 1995). Polyethylene microparticles of different sizes and charges can be separated by competition between electrostatic forces and acoustic field radiation forces (Yasuda et al, 1996A, B). In addition, ion permeation (for erythrocytes, Yasuda et al, 1997) and antibody production (for hybridoma cells, Pui. et al, 1995) were observed when mammalian cells were manipulated by the force of acoustic field radiation. During manipulation, no or minimal damage is done to the cells.

A standing wave plane can be generated in an acoustic field-field flow separation reaction cell (or acoustic field-electrophoresis-field flow separation, acoustic field-dielectrophoresis-field flow separation reaction cell) by applying an AC signal to the piezoelectric sensor. Similarly, by applying an AC signal on the piezoelectric sensor, a reaction cell with standing The sound field of wave components. Let us now examine the acoustic field radiation force experienced by particles in a standing wave field. Assume that the standing wave is established in a specific direction in the liquid (for example, the Z direction). The change of the standing wave in the Z direction can be expressed as follows:

Δp(z)=p ₀ sin(kz)cos(ωt)

Where Δp represents the sound field pressure in the Z direction, p ₀ represents the sound field pressure intensity, k represents the wave number (2π/λ, where λ is the wavelength), z represents the distance to the pressure node, ω represents the angular frequency, and t represents time. According to the theory proposed by Yoshioka and Kawashima in 1955, in a static standing wave field, the sound field radiation force acting on a spherical particle can be expressed by the following formula (see K.Yoshioka and Y.Kawashima in Acustica, 1955, Vol. 5, pages 167-173: Acoustic radiation pressure on a compressible sphere):

${\mathrm{<span\; class="notranslate">\; f</span>}}_{\mathrm{<span\; class="notranslate">\; acoustic</span>}}<span\; class="notranslate">\; =</span><span\; class="notranslate">\; -</span>\frac{\mathrm{<span\; class="notranslate">\; 4</span>}\mathrm{<span\; class="notranslate">\; \&pi;</span>}}{\mathrm{<span\; class="notranslate">\; 3</span>}}{\mathrm{<span\; class="notranslate">\; r</span>}}^{\mathrm{<span\; class="notranslate">\; 3</span>}}\mathrm{<span\; class="notranslate">\; k</span>}{\mathrm{<span\; class="notranslate">\; E.</span>}}_{\mathrm{<span\; class="notranslate">\; acoustic</span>}}\mathrm{<span\; class="notranslate">\; A</span>}\mathrm{<span\; class="notranslate">\; sin</span>}<span\; class="notranslate">\; (</span>\mathrm{<span\; class="notranslate">\; 2</span>}\mathrm{<span\; class="notranslate">\; kz</span>}<span\; class="notranslate">\; )</span>$

Where r is the radius of the particle, E _acoustic is the average energy density of the sound field, A is a constant, expressed by the following formula:

$\mathrm{<span\; class="notranslate">\; A</span>}<span\; class="notranslate">\; =</span>\frac{{\mathrm{<span\; class="notranslate">\; 5</span>}\mathrm{<span\; class="notranslate">\; \&rho;</span>}}_{\mathrm{<span\; class="notranslate">\; p</span>}}<span\; class="notranslate">\; -</span>{\mathrm{<span\; class="notranslate">\; 2</span>}\mathrm{<span\; class="notranslate">\; \&rho;</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}}{{\mathrm{<span\; class="notranslate">\; 2</span>}\mathrm{<span\; class="notranslate">\; \&rho;</span>}}_{\mathrm{<span\; class="notranslate">\; p</span>}}<span\; class="notranslate">\; +</span>{\mathrm{<span\; class="notranslate">\; \&rho;</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}}$ $\frac{{\mathrm{<span\; class="notranslate">\; \&gamma;</span>}}_{\mathrm{<span\; class="notranslate">\; p</span>}}}{{\mathrm{<span\; class="notranslate">\; \&gamma;</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}}$

Among them, ρ _m and ρ _p represent the density of particles and medium, and γ _m and γ _p represent the acoustic impedance of particles and medium. A here represents the polarization parameter of the sound field. The acoustic impedance is defined as the product of the density of the material (ρ _m and ρ _p represent the density of particles and media) and the sound velocity (C _m and C _p represent the sound speed in media and particles) (γ _m = ρ _m · C _m and γ _p = ρ _p · C _p ).

When A>0, the particle moves to the pressure node (z=0) of the standing wave.

When A<0, the particle moves away from the pressure node (z=0) of the standing wave.

Obviously, when particles with different densities and acoustic impedances are placed in the standing wave field, they are subject to different acoustic field radiation forces. For example: According to the established sound field energy distribution, the sound field radiation force experienced by a particle with a diameter of 10 microns varies from 0.01 to 1000pN.

In K.Yoshioka and Y.Kawashima in Acustica, 1955, Vol.5, pages 167-173: "Acoustic radiation pressure on a compressible sphere", the sound field radiation force that particles are subjected to in the traveling wave sound field is also described. Using these theories, experts who use sound fields to manipulate particles can analyze and calculate the sound field radiation force on particles.

Piezoelectric sensors are made of piezoelectric materials. When deformed by an applied mechanical force from an external piezoelectric material, the piezoelectric material generates an electric field (this is called the piezoelectric effect or dynamo effect). Conversely, applying an electric field to a piezoelectric material produces a mechanical stress (this is called the electrostriction or motor effect). The piezoelectric effect was discovered in 1880 by Pierre Curie and his brother Jacques. The reason for the explanation is that the displacement of ions leads to the electrical polarization of the structural units of the material. When an electric field is applied, the ions are displaced by electrostatic forces, resulting in mechanical denaturation of the entire material. In this way, in an acoustic field-field-flow separation or acoustic field-electrophoresis-field-flow separation or acoustic field-dielectrophoresis-field-flow separation device, an AC signal is applied to the piezoelectric sensor, and the vibration generated on the sensor is coupled into the reaction pool in the liquid, causing sound waves to be generated in the reaction cell. Such sound waves may have traveling and standing wave components.

Previously, acoustic waves have been used to separate particles in a medium. For example: US Patent No. 4523682 introduces a method for separating particles of different sizes, densities and properties in an acoustic wave reaction cell. The entirety of US Patent No. 4,523,682 is incorporated herein by reference. US Patent No. 4523682 describes the spatial separation of particles of different nature in acoustic waves. The characteristic of the device and method utilizing the acoustic wave-field flow separation of the present invention is that particles can be separated from mixtures and pure particle samples can be obtained. However, in US Patent No. 4,523,682, the particles can only be separated by the position of the particles in the acoustic wave reaction cell.

D.2 Electrophoretic force

中，对微粒的静电力F_E可以用下式表示：An electric field applied outside

In , the electrostatic force F _E on particles can be expressed by the following formula:

where _Qp represents the effective charge on the particle. The charge polarity of the particle and the direction of the applied electric field determine the direction of the electrostatic force on the particle.

D.3 Dielectrophoretic force

In a non-uniform electric field, the dielectrophoretic force F _DEPz on a particle with a radius r can be expressed by the following formula:

${\mathrm{<span\; class="notranslate">\; f</span>}}_{\mathrm{<span\; class="notranslate">\; DEPz</span>}}<span\; class="notranslate">\; =</span>{\mathrm{<span\; class="notranslate">\; 2</span>}\mathrm{<span\; class="notranslate">\; \&pi;\&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}{\mathrm{<span\; class="notranslate">\; r</span>}}^{\mathrm{<span\; class="notranslate">\; 3</span>}}{\mathrm{<span\; class="notranslate">\; \&chi;</span>}}_{\mathrm{<span\; class="notranslate">\; DEP</span>}}{<span\; class="notranslate">\; \&dtri;</span>\mathrm{<span\; class="notranslate">\; \&Egr;</span>}}_{\mathrm{<span\; class="notranslate">\; rms</span>}}^{\mathrm{<span\; class="notranslate">\; 2</span>}}<span\; class="notranslate">\; \&Center\; Dot;</span>{\stackrel{<span\; class="notranslate">\; \&Right\; Arrow;</span>}{\mathrm{<span\; class="notranslate">\; a</span>}}}_{\mathrm{<span\; class="notranslate">\; z</span>}}$

Where E _rms is the root mean square value of the electric field intensity, ε _m is the dielectric permeability of the medium, and χ _DEP is the polarization parameter of the particle, which is expressed by the following formula:

是复式通透率，(对于微粒x＝p，对于介质x＝m)。参数ε_p和σ_p是微粒的有效通透率和有效电导率。这些参数是由频率决定的。例如，一个细胞由于细胞膜的极化，有由频率决定的有效电导率和有效通透率。"Re" represents the real part of a complex number. symbol

is the multiplex permeability, (x=p for particles, x=m for media). The parameters _εp and _σp are the effective permeability and effective conductivity of the particle. These parameters are frequency dependent. For example, a cell has effective conductivity and effective permeability that depend on frequency due to the polarization of the cell membrane.

The above equation for the DEP force can also be written as:

${\mathrm{<span\; class="notranslate">\; f</span>}}_{\mathrm{<span\; class="notranslate">\; DEPz</span>}}<span\; class="notranslate">\; =</span>{\mathrm{<span\; class="notranslate">\; 2</span>}\mathrm{<span\; class="notranslate">\; \&pi;\&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}{\mathrm{<span\; class="notranslate">\; r</span>}}^{\mathrm{<span\; class="notranslate">\; 3</span>}}{\mathrm{<span\; class="notranslate">\; \&chi;</span>}}_{\mathrm{<span\; class="notranslate">\; DEP</span>}}{\mathrm{<span\; class="notranslate">\; V</span>}}^{\mathrm{<span\; class="notranslate">\; 2</span>}}\mathrm{<span\; class="notranslate">\; p</span>}<span\; class="notranslate">\; (</span>\mathrm{<span\; class="notranslate">\; z</span>}<span\; class="notranslate">\; )</span>{\stackrel{<span\; class="notranslate">\; \&Right\; Arrow;</span>}{\mathrm{<span\; class="notranslate">\; a</span>}}}_{\mathrm{<span\; class="notranslate">\; z</span>}}$

Wherein p(z) is a square wave field distribution of a unit pulse voltage (V=1V) on the electrode.

In an acoustic field-dielectrophoresis-field flow separation device, a non-uniform electric field can be generated by applying an AC signal to the microelectrode on the surface of the reaction cell. For a staggered electrode array, the dielectric force decays roughly exponentially in the direction perpendicular to the plane of the electrodes. This can be derived from Huang et al, 1997.

In the vertical direction of an acoustic field-dielectrophoresis-field flow separation reaction cell, particles or cells with different dielectric properties (ie, permeability and conductivity) will be subjected to different dielectric forces. For particles (including cells) manipulated by DEP, the dielectric force acting on a particle with a diameter of 10 microns varies between 0.01 and 1000 pN.

In the previously reported dielectrophoresis-field flow separation technique (Huang et al, Biophys.J.Vol.73, p1118-1129, 1997.; Wang et al., Biophys.J.Vol.74, p2689-2701, 1998.), only negative dielectrophoresis was used when influencing/controlling/manipulating the position of particles in the fluid velocity distribution. This is because forward dielectrophoresis may cause the particles to be adsorbed or captured to the electrode surface. However, in the acoustic field-dielectrophoresis-field flow separation method or device of the present invention, both positive dielectrophoresis and negative dielectrophoresis can be used. . Because the particles can be controlled into the fluid by adding a spatially variable sound field radiation force. Here, forward dielectrophoresis can be balanced by the spatially varying acoustic field radiation force.

D.4 Fluid lift

In an acoustic field-field flow separation reaction cell (or acoustic field-electrophoresis-field flow separation reaction cell or acoustic field-dielectrophoresis-field flow separation reaction cell) for particle separation and analysis, a fluid velocity distribution can be formed. In a rectangular reaction cell whose length and width are much greater than its height, a parabolic laminar fluid distribution can be formed. Such a velocity distribution can be expressed by:

${\mathrm{<span\; class="notranslate">\; V</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}<span\; class="notranslate">\; =</span>\mathrm{<span\; class="notranslate">\; 6</span>}<span\; class="notranslate">\; <</span>{\mathrm{<span\; class="notranslate">\; V</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}<span\; class="notranslate">\; ></span>\frac{\mathrm{<span\; class="notranslate">\; z</span>}}{\mathrm{<span\; class="notranslate">\; h</span>}}<span\; class="notranslate">\; (</span>\mathrm{<span\; class="notranslate">\; 1</span>}<span\; class="notranslate">\; -</span>\frac{\mathrm{<span\; class="notranslate">\; z</span>}}{\mathrm{<span\; class="notranslate">\; h</span>}}<span\; class="notranslate">\; )</span>$

where _Vm is the velocity of the fluid at z-height from the bottom of the reaction cell. H is the height of the reaction cell, <V _m > is the average flow rate of the fluid in the reaction cell. If a particle is transported by a fluid with such a velocity distribution, and the particle is located very close to the bottom or top of the reaction cell, it will experience a fluid lift force F _lift (the reaction cell is placed horizontally or nearly horizontally). If the distance between a particle and the bottom of the cell is very small (for example, a 10-µm particle is less than 1 µm from the bottom in a cell with a height of 200 µm), fluid lift can cause the particle to move away from the cell wall (Williams et al., 1992; 1994; 1996; 1997). This force is used in the classical supermembrane-field flow separation technique. Due to the balance of fluid lift and sink forces, particles are placed at different heights in the reaction tank (eg, Ratanathanawongs SK and Giddings, 1992, Williams et al, 1996). It is well known that when the distance between the particle and the pool wall increases, the fluid lift force decays to zero, but at a shorter distance, its influence should still be considered.

D.5 The balance of forces in the acoustic field-field flow separation, sound field-electrophoresis-field flow separation, and sound field-dielectric-field flow separation reaction cell

In a horizontal or nearly horizontal acoustic field-field flow separation reaction cell, the fluid flows along the horizontal direction. In the vertical direction, the particles are subjected to three forces, namely F _acoustic , F _lift and settling force $f_G=-\frac{4}{3}{\mathrm{\&pi;r}}^3({\mathrm{\&rho;}}_p-{\mathrm{\&rho;}}_m).$ When these forces balance:

F _acoustic (z)+F _lift (z)+F _G ＝0

If the force on the particle is zero, the particle will stop moving up and down. Due to the sound field force and fluid lift force determined by the height, the particle is balanced at the position h _eq where the force is zero. This position is determined by the energy distribution of the sound field, but more importantly by the particle density, acoustic impedance and particle size. Particles with different properties (density, acoustic impedance, size) will balance at different heights. And move at different speeds in the fluid (V _p , determined by _heq ). Therefore, the time of flow through the reaction cell is different (t _p =L/V _p ). Similar analysis can also be used for the acoustic field-field flow separation reaction cell which is not placed horizontally. In this case, the settling force effect on the particles in the fluid velocity distribution is different from that described above. Only the settling force component has an influence on the particle's position in the velocity distribution. Because only the component of the 90-degree angle between the direction of the sedimentation force and the direction of the flow movement works.

In a horizontal or nearly horizontal acoustic field-electrophoresis-field flow separation reaction cell, the fluid flows along the horizontal direction. In the vertical direction, the particles are subjected to four forces, namely F _acoustic , F _E , F _lift and the settling force $f_G=-\frac{4}{3}({\mathrm{\&rho;}}_p-{\mathrm{\&rho;}}_m).$ When these forces balance:

F _acoustic (z)+F _E +F _lift (z)+F _G ＝0

If the force on the particle is zero, the particle will stop moving up and down. Due to the sound field force and fluid lift force determined by the height, the particle is balanced at the position h _eq where the force is zero. This position is determined by the energy distribution of the acoustic field, the strength of the applied electric field, but more importantly by the particle density, acoustic impedance, and particle size. Particles with different properties (density, acoustic impedance, size) will balance at different heights. And move at different speeds in the fluid (V _p , determined by _heq ). Therefore, the time of flow through the reaction cell is different (t _p =L/V _p ). Compared with electrophoresis-field flow splitting, the acoustic field radiation force is added to the force balance equation. Similar analysis can also be used for acoustic field-electrophoresis-field flow separation reaction cells that are not placed horizontally. In this case, the settling force effect on the particles in the fluid velocity distribution is different from that described above. Only the settling force component has an influence on the particle's position in the velocity distribution. Because only the component of the 90-degree angle between the direction of the sedimentation force and the direction of the flow movement works.

By comparing the radiation force of the sound field with the electrostatic force (electrophoretic force), Yasuda et al. (1996A, B) proved that in a reaction cell with a height of 750 microns, polyethylene particles of different sizes and charges can be separated by a distance of 20 hours. Microns.

Similarly, in a horizontal or approximately horizontal acoustic field-dielectric field flow separation reaction cell, the fluid flows along the horizontal direction. The force balance equations of particles in the acoustic field-dielectric-field flow separation reaction cell are:

F _acoustic (z)+F _DEP (z)+F _lift (z)+F _G ＝0

Similarly, the position where the force on the particle is zero is the equilibrium height h _eq . This position is determined by the energy distribution of the acoustic field, the strength of the applied non-uniform electric field, but more importantly by the particle density, acoustic impedance, and particle size. Particles with different properties (density, acoustic impedance, size) will balance at different heights. And move at different speeds in the fluid (V _p , determined by _heq ). Therefore, the time of flow through the reaction cell is different (t _p =L/V _p ). Compared with dielectrophoresis-field flow splitting, the acoustic field radiation force is added to the force balance equation. Similar analysis can also be used for acoustic field-electrophoresis-field flow separation reaction cells that are not placed horizontally. Similar analysis can also be used for acoustic field-dielectric-field flow separation reaction cells that are not placed horizontally. In this case, the settling force effect on the particles in the fluid velocity distribution is different from that described above. Only the settling force component has an influence on the particle's position in the velocity distribution. Because only the component of the 90-degree angle between the direction of the sedimentation force and the direction of the flow movement works.

The above analysis is based on a single (large) particle, and the diffusion effect of the particle is ignored. Acoustic field-field flow separation, acoustic field-electrophoresis-field flow separation, and acoustic field-dielectric-field flow separation methods can also use the operation of small particles (down to the molecular level) for the separation and description of particles. A refinement of the above analysis should take into account diffusion effects. For example, Yasuda et al. (1996A) considered the diffusion effect of particles in the distribution of particle concentration in one-dimensional ultrasonic standing wave field and electric field. In these cases where particle diffusion effects are considered, the concentration distribution of particles in the vertical direction determines the elution time of a particular type of particle. In the vertical direction of the fluid, the theoretical analysis of the concentration distribution of small particles under the effect of field flow separation can be seen in many works on field flow separation (for example: Caldwell and Gao, 1993, Giddings 1993). Those experts who use field flow separation technology to separate and analyze molecules and small particles, as well as experts who use acoustic field/electrophoresis/dielectrophoretic effects to separate particles, will use acoustic field-field flow separation, acoustic field-electrophoresis-field flow separation , Acoustic field-dielectric-field flow separation is used for the separation and analysis of molecules and small particles for theoretical analysis.

Claims (77)

1. A flow point on the scene from utilize the device of sound field power recognition material, this device comprises:

   A) reaction tank comprises at least one ingress port and at least one outlet port, described reaction tank has such design feature, promptly when mounting medium was induced by this reaction tank, the translational speed of the described mounting medium of diverse location was different in reaction tank;

   B) have at least a piezoelectric sensor to form the part of described reaction tank, wherein said piezoelectric sensor can excite and produce sound wave by at least one road electric signal that is produced by electric signal generator, thereby cause at least one to put on the sound field power on the material in the described mounting medium, this sound field power has the component perpendicular to the mounting medium moving direction.
2. 2. according to the device described in the claim 1, comprise a plurality of ingress ports.
3. 3. according to the device described in the claim 1, comprise a plurality of outlet ports.
4. 4. according to the device described in the claim 1, wherein said outlet port is connected with gathering-device or identification apparatus.
5. 5. according to the device described in the claim 1, the length that the design feature of wherein said reaction tank is defined as reaction tank is fully greater than the height and the width of reaction tank.
6. 6. according to the device described in the claim 1, also comprise a plurality of piezoelectric sensors.
7. 7. according to the device described in the claim 6, wherein said a plurality of piezoelectric sensors are to excite by identical or different electric signal.
8. 8. according to the device described in the claim 6, wherein said a plurality of piezoelectric sensors are to be contained on the inside surface or outside surface of reaction tank.
9. 9. according to the device described in the claim 6, wherein said a plurality of piezoelectric sensors be arranged in one with mounting medium by on the fully parallel plane of the moving direction of reaction tank.
10. 10. according to the device described in the claim 1, the wherein said electric signal generator that is used to excite piezoelectric sensor to produce sound field power can be exported the electric signal of different amplitudes and frequency.
11. 11. according to the device described in the claim 1, wherein said reaction tank comprises a pipe.
12. 12. according to the device described in the claim 11, wherein said one or more piezoelectric sensors are to be distributed on the inside surface of pipe.
13. 13. according to the device described in the claim 11, wherein said piezoelectric sensor, or most of piezoelectric sensors wherein are to be distributed on the outside surface of pipe.
14. 14. according to the device described in the claim 1, wherein said reaction tank comprises a top cover, a bottom surface and two sidewalls.
15. 15. according to the device described in the claim 14, the velocity distribution of wherein said mounting medium diverse location in reaction tank is para-curve or approximate parabolical distribution.
16. 16. according to the device described in the claim 14, a wherein said piezoelectric sensor or a plurality of piezoelectric sensor are to be arranged on the reaction tank top cover.
17. 17. according to the device described in the claim 14, a wherein said piezoelectric sensor or a plurality of piezoelectric sensor are to be arranged on the reaction tank bottom surface.
18. 18. according to the device described in the claim 14, a wherein said piezoelectric sensor or a plurality of piezoelectric sensor are to be arranged on the interior facing surfaces of reaction tank.
19. 19. according to the device described in the claim 14, the height between wherein said reaction tank top and the bottom is the half-wavelength of sound wave standing wave.
20. 20. according to the device described in the claim 14, wherein said two parts that sidewall is the packing ring between top wall and base wall, and on packing ring, have passage.
21. 21. according to the device described in the claim 20, wherein said passage be shaped as rectangle, ellipse or circular.
22. 22. according to the device described in the claim 20, wherein said width of channel is between 1 millimeter to 20 centimetres, the height of passage is between 20 microns to 10 millimeters, and the length of passage is between 1 centimetre to 200 centimetres.
23. 23. a flow point on the scene from utilize the device of sound field power recognition material, this device comprises:

    A) reaction tank comprises at least one ingress port and at least one outlet port, described reaction tank has such design feature, promptly when mounting medium was induced by this reaction tank, the translational speed of the described mounting medium of diverse location was different in reaction tank;

    B) have at least a piezoelectric sensor to be positioned at certain part of described reaction tank, wherein said piezoelectric sensor can excite the generation sound wave by at least one road electric signal that is produced by electric signal generator, thereby cause at least one to put on the sound field power on the material in the described mounting medium, this sound field power has the component perpendicular to the mounting medium moving direction.
24. 24. a flow point on the scene from utilize the device of sound field power recognition material, this device is by constituting with lower member:

    A) reaction tank comprises at least one ingress port and at least one outlet port, described reaction tank has such design feature, promptly when mounting medium was induced by this reaction tank, the translational speed of the described mounting medium of diverse location was different in reaction tank;

    B) have at least a piezoelectric sensor to form the part of described reaction tank, wherein said piezoelectric sensor can excite the generation sound wave by at least one road electric signal that is produced by electric signal generator, thereby cause at least one to put on the sound field power on the material in the described mounting medium, this sound field power has the component perpendicular to the mounting medium moving direction.
25. 25. a flow point on the scene from utilize the device of electrophoretic force harmony field force recognition material, this device is by constituting with lower member:

    A) reaction tank comprises at least one ingress port and at least one outlet port, described reaction tank has such design feature, promptly when mounting medium was induced by this reaction tank, the translational speed of the described mounting medium of diverse location was different in reaction tank;

    B) have at least two electrodes to form the part of described reaction tank, wherein said electrode can excite the generation electric field by at least one road electric signal that is produced by electric signal generator, thereby cause at least one to put on the electrophoretic force on the material in the described mounting medium, this electrophoretic force has the component perpendicular to the mounting medium moving direction;

    C) have at least a piezoelectric sensor to form the part of described reaction tank, wherein said piezoelectric sensor can excite the generation sound wave by at least one road electric signal that is produced by electric signal generator, thereby cause at least one to put on the sound field power on the material in the described mounting medium, this sound field power has the component perpendicular to the mounting medium moving direction.
26. 26., comprise plural electrode according to the device described in the claim 25.
27. 27. according to the device described in the claim 25, each electrode in the wherein said plural electrode all is connected on the electrical lead in the electrical lead bus that is electrically connected with electric signal generator individually.
28. 28. according to the device described in the claim 25, wherein said electrode is to be distributed on the reaction tank vertical or horizontally.
29. 29. according to the device described in the claim 25, wherein said distribution of electrodes is on the inside surface of reaction tank.
30. 30. according to the device described in the claim 25, wherein said electrode be arranged in one with the fully parallel plane of the moving direction of mounting medium by reaction tank on.
31. 31. according to the device described in the claim 25, wherein said electrode forms an electrod-array, described electrod-array can be a mutual dislocation formula electrod-array, mutual dislocation formula castle electrod-array has the mutual dislocation formula electrod-array of cycle apex and has the mutual dislocation formula electrod-array on cycle circular arc top.
32. 32. according to the device described in the claim 25, wherein said electrode is a metal level that is deposited on the reaction tank surface.
33. 33. according to the device described in the claim 32, wherein said metal can be a gold, platinum, aluminium, chromium, titanium, copper and silver.
34. 34. according to the device described in the claim 25, the wherein said electric signal generator that is used for exciting electrode generation electrophoretic force is one and can exports the galvanic direct voltage source of different amplitudes or the alternating-current voltage source that can export different amplitudes, phase place.
35. 35. according to the device described in the claim 25, the wherein said electric signal that is used for exciting electrode generation electrophoretic force is the ac signal of a direct current electric signal or a low frequency.
36. 36. according to the device described in the claim 25, wherein said reaction tank comprises a pipe.
37. 37. according to the device described in the claim 36, a wherein said electrode and/or piezoelectric sensor, or a plurality of electrode and/or piezoelectric sensor are to be distributed on the inside surface of pipe.
38. 38. according to the device described in the claim 36, a wherein said piezoelectric sensor, or a plurality of piezoelectric sensor is to be distributed on the outside surface of pipe.
39. 39. according to the device described in the claim 25, wherein said reaction tank comprises a top cover, a bottom surface and two sidewalls, and a described electrode and/or piezoelectric sensor, or a plurality of electrode and/or piezoelectric sensor are to be distributed on the top cover of reaction tank.
40. 40. according to the device described in the claim 25, wherein said reaction tank comprises a top cover, a bottom surface and two sidewalls, and a described electrode and/or piezoelectric sensor, or a plurality of electrode and/or piezoelectric sensor are to be distributed on the bottom surface of reaction tank.
41. 41. according to the device described in the claim 25, a wherein said electrode and/or piezoelectric sensor, or a plurality of electrode and/or piezoelectric sensor are to be arranged in the reaction tank on the facing surfaces.
42. 42. a flow point on the scene from utilize the device of electrophoretic force harmony field force recognition material, this device comprises:

    A) reaction tank comprises at least one ingress port and at least one outlet port, described reaction tank has such design feature, promptly when mounting medium was induced by this reaction tank, the translational speed of the described mounting medium of diverse location was different in reaction tank;

    B) have at least two electrodes to form the part of described reaction tank, wherein said electrode can excite the generation electric field by at least one road electric signal that is produced by electric signal generator, thereby cause at least one to put on the electrophoretic force on the material in the described mounting medium, this electrophoretic force has the component perpendicular to the mounting medium moving direction;

    C) have at least a piezoelectric sensor to form the part of described reaction tank, wherein said piezoelectric sensor can excite the generation sound wave by at least one road electric signal that is produced by electric signal generator, thereby cause at least one to put on the sound field power on the material in the described mounting medium, this sound field power has the component perpendicular to the mounting medium moving direction.
43. 43. a flow point on the scene from utilize the device of electrophoretic force harmony field force recognition material, this device to have to constitute with lower member:

    A) reaction tank comprises at least one ingress port and at least one outlet port, described reaction tank has such design feature, promptly when mounting medium was induced by this reaction tank, the translational speed of the described mounting medium of diverse location was different in reaction tank;

    B) have at least two electrodes to form the part of described reaction tank, wherein said electrode can excite the generation electric field by at least one road electric signal that is produced by electric signal generator, thereby cause at least one to put on the electrophoretic force on the material in the described mounting medium, this electrophoretic force has the component perpendicular to the mounting medium moving direction;

    C) have at least a piezoelectric sensor to form the part of described reaction tank, wherein said piezoelectric sensor can excite the generation sound wave by at least one road electric signal that is produced by electric signal generator, thereby cause at least one to put on the sound field power on the material in the described mounting medium, this sound field power has the component perpendicular to the mounting medium moving direction.
44. 44. a flow point on the scene from utilize the device of dielectrophoresis force harmony field force recognition material, this device is by constituting with lower member:

    A) reaction tank comprises at least one ingress port and at least one outlet port, described reaction tank has such design feature, promptly when mounting medium was induced by this reaction tank, the translational speed of the described mounting medium of diverse location was different in reaction tank;

    B) have at least two electrodes to form the part of described reaction tank, wherein said electrode can excite by at least one road electric signal that is produced by electric signal generator and produce a non-uniform electric field, thereby cause at least one to put on the dielectrophoresis force on the material in the described mounting medium, this dielectrophoresis force has a component perpendicular to the mounting medium moving direction;

    C) have at least a piezoelectric sensor to form the part of described reaction tank, wherein said piezoelectric sensor can excite the generation sound wave by at least one road electric signal that is produced by electric signal generator, thereby cause at least one to put on the sound field power on the material in the described mounting medium, this sound field power has the component perpendicular to the mounting medium moving direction.
45. 45., comprise plural electrode according to the device described in the claim 44.
46. 46. according to the device described in the claim 45, each electrode in the wherein said plural electrode all is connected on the electrical lead in the electrical lead bus that is electrically connected with electric signal generator individually.
47. 47. according to the device described in the claim 45, wherein said electrode further can also produce the space non-uniform electric field.
48. 48. according to the device described in the claim 44, the electric signal that the wherein said electric signal generator output that is used for exciting electrode generation dielectrophoresis force has different amplitudes and phase place.
49. 49. a flow point on the scene from utilize the device of dielectrophoresis force harmony field force recognition material, this device comprises:

    A) reaction tank comprises at least one ingress port and at least one outlet port, described reaction tank has such design feature, promptly when mounting medium was induced by this reaction tank, the translational speed of the described mounting medium of diverse location was different in reaction tank;

    B) have at least two electrodes to form the part of described reaction tank, wherein said electrode can excite by at least one road electric signal that is produced by electric signal generator and produce a non-uniform electric field, thereby cause at least one to put on the dielectrophoresis force on the material in the described mounting medium, this dielectrophoresis force has a component perpendicular to the mounting medium moving direction;

    C) have at least a piezoelectric sensor to form the part of described reaction tank, wherein said piezoelectric sensor can excite the generation sound wave by at least one road electric signal that is produced by electric signal generator, thereby cause at least one to put on the sound field power on the material in the described mounting medium, this sound field power has the component perpendicular to the mounting medium moving direction.
50. 50. a flow point on the scene from utilize the method for sound field power recognition material, this method comprises:

    A) obtain the device described in the claim 1;

    B) mounting medium that will contain material to be identified of the ingress port by device described in the claim 1 imports in the reaction tank of device, and wherein said importing makes mounting medium pass through reaction tank according to velocity distribution;

    C) excite the generation sound wave by piezoelectric sensor being applied at least one road electric signal that produces by electric signal generator, thereby cause at least one to put on the sound field power on the material in the described mounting medium, this sound field power has the component perpendicular to the mounting medium moving direction;

    Be in described material in the described mounting medium in certain position that is shifted onto under the effect of power on the direction vertical, come recognition material according to the difference of material distributing position on this direction with the mounting medium moving direction.
51. 51. according to the method described in the claim 50, wherein said identification to material is to realize by the velocity distribution of mounting medium by reaction tank, the speed dependent that material moves in reaction tank is in its residing position in velocity distribution.
52. 52. according to the method described in the claim 50, it is to depend on their positions velocity distribution that wherein said material moves out the time interval of coming from the outlet port of reaction tank.
53. 53. according to the method described in the claim 50, it is to depend on their positions in velocity distribution that the outlet port of wherein said material from a plurality of outlet ports of reaction tank moves out the time interval of coming.
54. 54., wherein act on gravity direction on the material perpendicular to the moving direction of mounting medium by reaction tank according to the method described in the claim 50.
55. 55. according to the method described in the claim 50, wherein said material to be identified comprises cell, organelle, virus, molecule or their condensate or complex.
56. 56. according to the method described in the claim 55, wherein said cell comprises zooblast, vegetable cell, mushroom cell, bacterial cell, cultured cell or recombinant cell.
57. 57. according to the method described in the claim 55, wherein said organelle comprises nucleus, mitochondria, chloroplast, ribosomes, thick matter endoplasmic reticulum, golgiosome, lysosome, proteasome, excretion vesicles, vacuole or microsome.
58. 58. according to the method described in the claim 55, wherein said molecule comprises inorganic molecule, organic molecule or their complex.
59. 59. according to the method described in the claim 58, wherein said inorganic molecule comprises sodium, potassium, magnesium, calcium, chlorine, iron, copper, zinc, manganese, cobalt, iodine, molybdenum, vanadium, nickel, chromium, fluorine, silicon, tin, boron or arsenic ion.
60. 60. according to the described method of claim 58, wherein said organic molecule comprises amino acid, peptide, protein, nucleosides, nucleotide, oligonucleotides, nucleic acid, vitamin, monose, oligosaccharides, carbohydrates, lipid or their compound.
61. 61. according to the described method of claim 50, the size of wherein said material to be identified is between 0.01 micron to 1000 microns.
62. 62. a flow point on the scene from utilize the method for sound field power recognition material, this method comprises:

    A) obtain the device described in the claim 1;

    B) ingress port by device described in the claim 1 imports to mounting medium in the reaction tank of device and is full of the entire reaction pond until mounting medium;

    C) sample that will contain material to be identified injects the interior mounting medium of reaction tank;

    D) piezoelectric sensor is applied at least one road electric signal that is produced by electric signal generator, the wherein said piezoelectric sensor that excites has produced sound wave, thereby causes having at least a sound field power to be applied on the material;

    E) ingress port by device imports to mounting medium in the reaction tank of device, and wherein said importing makes mounting medium flow through reaction tank according to a kind of velocity distribution;

    Be in described material in the described mounting medium in certain position that is shifted onto under the effect of power on the direction vertical, come recognition material according to the difference of material distributing position on this direction with the mounting medium moving direction.
63. 63. according to the method described in the claim 62, wherein saidly piezoelectric sensor is applied electric signal make its generation be applied to sound field power on the described material to have caused adding mounting medium and mounting medium is complied with before certain velocity distribution flows through reaction tank in reaction tank, the material in the mounting medium is shifted onto the equilibrium position on the direction vertical with the mounting medium moving direction.
64. 64. a flow point on the scene from utilize the method for electrophoretic force harmony field force recognition material, this method comprises:

    A) obtain the device described in the claim 25;

    B) ingress port by device described in the claim 25 imports to mounting medium in the reaction tank of device, and wherein said importing makes mounting medium pass through reaction tank according to velocity distribution;

    C) excite the generation electric field by electrode being applied at least one road electric signal that is produced by electric signal generator, thereby cause at least one to put on the electrophoretic force on the material in the described mounting medium, this electrophoretic force has the component perpendicular to the mounting medium moving direction;

    D) excite the generation sound wave by piezoelectric sensor being applied other at least one road electric signal that produces by electric signal generator, thereby cause at least one to put on the sound field power on the material in the described mounting medium, this sound field power has the component perpendicular to the mounting medium moving direction;

    Be in described material in the described mounting medium in certain position that is shifted onto under the effect of power on the direction vertical, come recognition material according to the difference of material distributing position on this direction with the mounting medium moving direction.
65. 65. according to the described method of claim 64, wherein said electrophoretic force harmony field force is produced simultaneously.
66. 66. according to the described method of claim 64, wherein said electrophoretic force harmony field force is that order produces.
67. 67. a flow point on the scene from utilize the method for electrophoretic force harmony field force recognition material, this method comprises:

    A) obtain the device described in the claim 25;

    B) ingress port by device described in the claim 25 imports to mounting medium in the reaction tank of device and is full of the entire reaction pond until mounting medium;

    C) sample that will contain material to be identified injects the interior mounting medium of reaction tank;

    D) electrode is applied at least one road electric signal that is produced by electric signal generator, wherein said exciting electrode has produced an electric field, thereby causes having at least an electrophoretic force to be applied on the described material;

    E) piezoelectric sensor is applied another road electric signal at least that is produced by electric signal generator, the wherein said piezoelectric sensor that excites has produced sound wave, thereby causes having at least a sound field power to be applied on the described material;

    F) ingress port by device imports to mounting medium in the reaction tank of device, and wherein said importing makes mounting medium pass through reaction tank according to velocity distribution;

    Be in described material in the described mounting medium in certain position that is shifted onto under the effect of power on the direction vertical, come recognition material according to the difference of material distributing position on this direction with the mounting medium moving direction.
68. 68. according to the method described in the claim 67, wherein saidly electrode is applied electric signal make it produce at least one electrophoretic force to be applied on the described material, piezoelectric sensor is applied electric signal to be made it produce at least one sound field power to be applied on the described material, caused in reaction tank, add mounting medium make mounting medium according to the velocity distribution form by before the reaction tank, the species distribution in the mounting medium perpendicular to mounting medium by the equilibrium position on the reaction tank direction.
69. 69. according to the described method of claim 67, wherein said electrophoretic force harmony field force is produced simultaneously.
70. 70. according to the described method of claim 67, wherein said electrophoretic force harmony field force is that order produces.
71. 71. a flow point on the scene from utilize the method for dielectrophoresis force harmony field force recognition material, this method comprises:

    A) obtain the device described in the claim 44;

    B) ingress port by device described in the claim 44 imports to mounting medium in the reaction tank of device, and wherein said importing makes mounting medium pass through reaction tank according to velocity distribution;

    C) electrode is applied at least one road electric signal that is produced by electric signal generator, wherein said exciting electrode has produced an inhomogeneous field, thereby causes having at least a dielectrophoresis force that is applied on the described material to have a component that moves through the reaction tank direction perpendicular to mounting medium;

    D) piezoelectric sensor is applied another road electric signal at least that is produced by electric signal generator, the wherein said piezoelectric sensor that excites has produced a sound wave, thereby cause at least one to put on the sound field power on the material in the described mounting medium, this sound field power has the component perpendicular to the mounting medium moving direction;

    Be in described material in the described mounting medium in certain position that is shifted onto under the effect of power on the direction vertical, come recognition material according to the difference of material distributing position on this direction with the mounting medium moving direction.
72. 72. according to the described method of claim 71, wherein said dielectrophoresis force harmony field force is produced simultaneously.
73. 73. according to the described method of claim 71, wherein said dielectrophoresis force harmony field force is that order produces.
74. 74. a flow point on the scene from utilize the method for dielectrophoresis force harmony field force recognition material, this method comprises:

    A) obtain the device described in the claim 44;

    B) ingress port by device described in the claim 44 imports to mounting medium in the reaction tank of device and is full of the entire reaction pond until mounting medium;

    C) sample that will contain material to be identified injects the interior mounting medium of reaction tank;

    D) electrode is applied at least one road electric signal that is produced by electric signal generator, wherein said exciting electrode has produced an electric field, thereby causes having at least a dielectrophoresis force to be applied on the described material;

    E) piezoelectric sensor is applied another road electric signal at least that is produced by electric signal generator, the wherein said piezoelectric sensor that excites has produced sound wave, thereby causes having at least a sound field power to be applied on the described material;

    F) ingress port by device imports to mounting medium in the reaction tank of device, and wherein said importing makes mounting medium pass through reaction tank according to velocity distribution;

    Be in described material in the described mounting medium in certain position that is shifted onto under the effect of power on the direction vertical, come recognition material according to the difference of material distributing position on this direction with the mounting medium moving direction.
75. 75. according to the method described in the claim 74, wherein saidly electrode is applied electric signal make it produce at least one dielectrophoresis force to be applied on the described material, piezoelectric sensor is applied electric signal to be made it produce at least one sound field power to be applied on the described material, caused adding mounting medium and mounting medium is complied with before certain velocity distribution flows through reaction tank in reaction tank, the material in the mounting medium is shifted onto the equilibrium position on the direction vertical with the mounting medium moving direction.
76. 76. according to the described method of claim 74, wherein said dielectrophoresis force harmony field force is produced simultaneously.
77. 77. according to the described method of claim 74, wherein said dielectrophoresis force harmony field force is that order produces.

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