EP1092144A1 - Procede et dispositif de manipulation de particules dans un microsysteme - Google Patents

Procede et dispositif de manipulation de particules dans un microsysteme

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Publication number
EP1092144A1
EP1092144A1 EP99931204A EP99931204A EP1092144A1 EP 1092144 A1 EP1092144 A1 EP 1092144A1 EP 99931204 A EP99931204 A EP 99931204A EP 99931204 A EP99931204 A EP 99931204A EP 1092144 A1 EP1092144 A1 EP 1092144A1
Authority
EP
European Patent Office
Prior art keywords
microsystem
particles
forces
channel
centrifuge
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP99931204A
Other languages
German (de)
English (en)
Inventor
Günter FUHR
Rolf Hagedorn
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Revvity Cellular Technologies GmbH
Original Assignee
Evotec Biosystems GmbH
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from DE1998153658 external-priority patent/DE19853658A1/de
Application filed by Evotec Biosystems GmbH filed Critical Evotec Biosystems GmbH
Publication of EP1092144A1 publication Critical patent/EP1092144A1/fr
Withdrawn legal-status Critical Current

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Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/50273—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the means or forces applied to move the fluids
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502761—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C5/00—Separating dispersed particles from liquids by electrostatic effect
    • B03C5/02—Separators
    • B03C5/022—Non-uniform field separators
    • B03C5/026—Non-uniform field separators using open-gradient differential dielectric separation, i.e. using electrodes of special shapes for non-uniform field creation, e.g. Fluid Integrated Circuit [FIC]
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
    • B01L2200/0652—Sorting or classification of particles or molecules
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0825—Test strips
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0409—Moving fluids with specific forces or mechanical means specific forces centrifugal forces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0415—Moving fluids with specific forces or mechanical means specific forces electrical forces, e.g. electrokinetic
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0415—Moving fluids with specific forces or mechanical means specific forces electrical forces, e.g. electrokinetic
    • B01L2400/0424—Dielectrophoretic forces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0457—Moving fluids with specific forces or mechanical means specific forces passive flow or gravitation
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S977/00—Nanotechnology
    • Y10S977/902—Specified use of nanostructure
    • Y10S977/904—Specified use of nanostructure for medical, immunological, body treatment, or diagnosis

Definitions

  • the invention relates to a method for manipulating particles in fluidic microsystems, in particular for moving particles in microsystems along predetermined, at least sectionally straight paths, and devices for implementing such a method, in particular a fluidic microsystem, in which synthetic or biological particles in a suspension liquid manipulated, and applications of such a microsystem.
  • Fluidic microsystems with structures through which liquids flow e.g. channels
  • microelectrodes for influencing particles e.g. biological cells
  • high-frequency fields are based on negative or positive dielecrophoresis
  • Fluidic microsystems are usually flowed through by a liquid for propelling the particles.
  • the microelectrodes applied on both sides of the channel (top, bottom) lead to a compartmentalization of the channel by means of high-frequency electrical fields, with which the suspended particles can be deflected in the desired manner, for example via branches in adjacent channels or other structural elements. Difficulties are caused especially by the flushing in of the particles at each end of the channel and the setting of the generally low flow velocities (a few ⁇ l / h), which always entail serious restrictions with increasing miniaturization.
  • a general disadvantage of conventional fluidic microsystems is that a solution flow is required for the directed and adjustable particle movement, the control of which (eg the flow rate) causes problems.
  • a central flow system in which liquid flows in a microsystem are not adjusted with conventional pumps and valves, but under the action of centrifugal forces.
  • the microsystem is located in a disk-shaped carrier in the form of a CD-ROM disk.
  • the carrier is intended to be rotated at high speed (in the range from 100 to 10,000 revolutions per minute).
  • the liquids in the microsystem move radially outwards under the action of the centrifugal forces.
  • certain biochemical reactions take place in the microsystem. It is also contemplated to use the liquid movement to transport particles, such as in a conventionally pumped liquid flow.
  • the centrifugal technology according to MJ Madou et al. has the following disadvantages. Both the achievement of a sufficient fluid movement as well as a possible handicap-free entrainment of particles with the liquid in the disc-shaped, plane rotor inevitably require the high speeds of the carrier mentioned. This results in a limitation of the conventional centrifugal flow system to certain basic functions of conventional centrifugation or the achievement of biochemical reactions.
  • the above-mentioned microelectrode technology for generating high-frequency electrical fields in the microstructures cannot be used.
  • Another disadvantage relates to the particle sorting and realized with conventional centrifugal technology Counts. These are only possible by producing microchannels with a size that corresponds to the size of the particles to be processed. This means that a given microsystem is always limited to a certain particle size. In addition, the handling of biological particles (cells, cell components) quickly leads to interactions between the particles and the channel wall, which lead to channel blockages.
  • Centrifuge systems are also generally known, in which the sample material in the centrifuge not only the centrifugal forces, but also additionally e.g. Magnetic or electrical forces are exposed to achieve specific separation effects depending on the ratio of centrifugal and additional forces.
  • these centrifuge systems cannot be used to manipulate biological objects.
  • Biological objects e.g. cells
  • Such conductivities would lead to undesirable heating effects in the conventional centrifuge systems with relatively large electrode areas.
  • the conventional centrifuge systems are therefore based on conductivities of approx. 0.1 Siemens / m limited.
  • the object of the invention is to provide an improved method for manipulating particles in fluidic microsystems, with which the disadvantages of conventional microsystems are overcome and which has an expanded area of application.
  • the object of the invention is also to provide an improved fluidic microsystem with a directional particle movement that is simplified and adjustable with high accuracy.
  • the object of the invention is also to specify applications of such an improved microsystem.
  • a first important aspect of the invention is to deviate from the conventional centrifugal flow system with moving liquids to a procedure in which only the particles to be manipulated are moved in a fluidic microsystem under the action of centrifugal forces, with essentially no liquid flows or movements in the microsystem occur.
  • a series of measures are implemented, which include in particular the use of a fluidic microsystem which is closed at least on one side, the attachment of such a microsystem to a vibrating rotor centrifuge device and the operation of this centrifuge device at a predetermined speed at which the particles in the microsystem are in the desired manner move.
  • the method according to the invention enables centrifugation processes at low speeds. Due to the use of an oscillating rotor system, in which a rotor as a support for the microsystem rises from a vertical orientation (at standstill or at low speeds) to a horizontal orientation (at high speeds), the gravitational force increasingly influences the movement of the particles as the speed decreases in the microsystem. According to a further aspect of the invention, a particle movement is also described in microsystems which are closed on at least one side and which are at a standstill with the microsystem oriented vertically. The particle movement takes place as sedimentation under the effect of gravitational force.
  • microsystems which are equipped with microelectrode devices for dielectrophoretically influencing the particle movement, are combined in particular with the principle of centrifuging. Due to the centrifugal forces, the suspended particles move through the microchannels or other microstructures in a microsystem, in which they are separated (without being able to escape) under the action of electrical polarization forces, for example, brought into a predetermined position, fused, sorted or permeated.
  • An important advantage of the invention is that, for the first time in complexly structured microsystems with dielectrophoretic particle influence, the use of difficult to control and fault-prone pumps or valves can be dispensed with, without the functionality of the microsystem being restricted. There are no restrictions on the cross channel dimensions. It is possible to set the microsystem in rotation simultaneously with the associated control electronics. Interactions of particles (in particular biological particles) with wall areas of the microsystem can easily be avoided or can also be achieved in a predetermined manner with the appropriate structuring for the investigation of binding processes.
  • An important advantage of the invention is that all particles are equally exposed to the centrifugal force and move along predetermined channels according to a reference direction and the separation z. B. is achieved in different subchannels or reservoirs exclusively via deflection forces, which act particle-specific regardless of the centrifugal force.
  • the deflection forces have a direction deviating from the reference direction, the angle difference preferably being less than 90 °. Only the particle speed is set via the centrifugal force. After the separation, the additional forces can be switched off without the particles mixing again. It is an unexpected and important feature that the use of a vibratory rotor centrifuge prevents the contact of particles with sample chamber walls, which is particularly important for biological objects.
  • FIG. 1 is a schematic perspective view of an inventive construction of a centrifuge with a microsystem
  • Fig. 2 is a schematic plan view of a microsystem according to the invention, which is set up for particle separation, and
  • FIG. 3 shows a schematic top view of a programmable loading microsystem according to a further embodiment of the invention.
  • the embodiments of the invention described here relate to the combination of a microsystem which is equipped with a microelectrode device for performing negative or positive dielectrophoresis (dielectrophoretic microsystem) with an oscillating rotor centrifuge device.
  • dielectrophoretic microsystem an oscillating rotor centrifuge device.
  • Both the dielectrophoretic microsystem (apart from the fact that channel structures can be closed at least on one side) and the vibratory rotor centrifuge device are known per se, so that their technical details are not discussed further here.
  • each centrifuge device direction is included with at least one speed-dependent erectable rotor, which itself forms the microsystem and the associated control, in which the microsystem and the associated control are integrated or on which the microsystem and the associated control are placed.
  • the particles manipulated according to the invention can comprise synthetic particles or biological objects.
  • the synthetic particles are, for example, membrane-encased structures, such as liposomes or vesicles, or so-called beads or also macromolecules.
  • the biological objects include, for example, biological cells or components thereof (e.g. cell organelles), bacteria or viruses.
  • the particles can also be aggregates or agglomerations of such particles and / or objects.
  • the invention is preferably implemented with fluids relevant to cell physiology or medicine with conductivities below 5 Siemens / m.
  • FIG. 1 is a schematic overview representation of a device according to the invention for illustrating the attachment of a dielectrophoretic system to a centrifuge device.
  • the control electronics are connected to the microsystem 15 via cables 14, plugs or otherwise.
  • the control device is preferably supplied with power via an electrical connection (all-round contact) with the fixed laboratory system.
  • the microsystem has an input depot 16 which, depending on the application, can be of different sizes can and is filled with a particle or cell suspension before centrifugation.
  • a channel structure extends from the input depot 16 to collecting zones 17a, 17b, which form an end of the microsystem 15 that is closed at least during centrifugation.
  • the microsystem 15 is arranged on the receptacle 12 in such a way that when the centrifuge device is operating (rotation of the rotor about the axis of rotation 11 at the rotation frequency ⁇ ), the centrifugal forces acting on the microsystem 15 and particles therein are in the reference direction from the input depot 18 to Collection zones 17a, 17b are directed.
  • the receptacles 12 are pivotally attached to the rotor (not shown). When the centrifuge is at a standstill, the receptacle 12 is oriented essentially vertically or at a slight angle with respect to the axis of rotation. In centrifuge operation, the receptacles 12 align themselves in a larger angle, depending on the speed, up to the horizontal orientation perpendicular to the axis of rotation 11. Under the effect of the gravitational force (when the centrifuge is stopped) or As a result of the centrifugal forces, the particles pass through the electronically controlled microchannel system and collect in the collecting zones (eg at the closed end of the part of the microsystem pointing away from the rotor axis).
  • the particles are treated according to predetermined programs (see below). Since the particles perform different movements depending on their density and assume end positions, the advantage of centrifugal separation and movement is combined with the possibilities of programmable dielectrophoresis in the present invention. As a rule, negative dielectrophoresis, in exceptional cases positive dielectrophoresis of the particles is also used.
  • a further advantage of the invention is the control of the particle movement via the rotational speed ( ⁇ ) of the rotor 11. Since programmable variations can also be run through here, a second complex of definable parameters for the particle manipulation is given.
  • the centrifuge device is provided with a speed control (not shown), which is set up for reproducible and precise speed setting, in particular in low speed ranges.
  • the speed is selected depending on the application, depending on the desired speed of the particles to be manipulated and depending on the specific centrifuge design.
  • the particle speeds of interest for biological particles (eg cells) are below approx. 500 ⁇ m / s (preferably in the range from 50 to 100 ⁇ m / s) and for synthetic particles (eg latex beads) at higher speeds (eg a few mm / s).
  • the speed of the centrifuge device is selected according to the relationship between speed and centrifugal force depending on the size or mass density of the particles.
  • the following information relates to a distance of the microsystem from the rotor axis in the range of 1 to 10 cm.
  • the speeds can be in the range from 1 to 1000 U / mm, for example.
  • particles with a diameter of approx. 5 ⁇ m speeds up to 100 U / mm are preferred, although higher speeds can also be set. With particularly small particles, for example macromolecules, even higher speeds can be achieved.
  • centrifugal forces that can be achieved are in the range from pN to nN.
  • the centrifuge device is however also designed for higher speeds, which can be set especially for small particles or for cleaning or winding purposes. These increased speeds can range up to the range of speeds of conventional laboratory centrifuges.
  • the speed of the centrifuge is also selected depending on the dielectrophoretic forces that act on the particles in the microsystem.
  • the dielectrophoretic forces are dependent on the type and size of the particles as polarization forces.
  • the speed is preferably selected so that the centrifugal forces on the particles are less than or equal to the dielectrophoretic forces. If these are not known, the speed can also be selected in relation to the following criterion.
  • the particles have to move so slowly through the channel structure that there is sufficient time for dielectrophoretic deflection when passing the microelectrode devices.
  • the effectiveness or ineffectiveness of the dielectrophoretic deflection as a function of the speed can be detected optically or electrically using suitable sensors.
  • Fig. 2 shows schematically a microsystem for the separation of a particle mixture, consisting of larger particles 21 (e.g. cells) and small particles 22, which are in a suspension.
  • the centrifugal forces act in the direction of arrow 23 (reference direction).
  • the typical dimensions of the channel structure 24 are as follows:
  • Width a few 10 ⁇ m to a few mm
  • Length a few mm to a few cm
  • microelectrodes 27a, 27b are arranged opposite one another, which when actuated with an alternating voltage (usually a frequency in the MHz range and an amplitude of a few volts) produce field barriers across the channel which are negative (also requires positive) dielectrophoresis to deflect the particles (in the case shown here, the large particles).
  • an alternating voltage usually a frequency in the MHz range and an amplitude of a few volts
  • the channel structure 24 extends from the input depot 28 to the closed channel ends 29a, 29b, into which the straight channel branches in a central section.
  • a first pair of microelectrodes 27a, 27b is arranged directly at the channel-side end of the input depot 28 to form a field barrier which projects obliquely into the channel and has the task of forcing the large particles 21 into the part of the channel 24 which is on the right in plan view.
  • a second pair of the microelectrodes 27a, 27b is arranged immediately before the branching to the channel ends 29a, 29b and forms a field barrier which extends obliquely across the channel width into the branch leading to the channel end 29b and is intended for the large particles 21 to this Lead end of channel.
  • a manipulation method according to the invention which in this example is aimed at separating the particles, comprises the following steps.
  • the microsystem Before the centrifugation, the microsystem is filled with a suitable liquid.
  • the microsystem is already installed in a receptacle 12 in the centrifuge (see FIG. 1). Installation can also take place after the microsystem has been filled.
  • the electrodes 27a, 27b are activated and the suspension of the particles to be separated is added to the input depot 28, for example with a pipetting device.
  • the centrifuge device is initially still at rest, ie the microsystem is vertical or slightly inclined to the vertical. The gravitational force that acts on the particles leads to a mass-dependent drop in the channel structure (sedimentation).
  • the further movement of the particles towards the channel ends takes place e depending on the desired particle speed exclusively under the effect of the gravitational force or under the joint effect of the gravitational force and the centrifugal forces.
  • the centrifugation can thus be understood as sedimentation under the effect of an artificially increased acceleration of gravity.
  • the moving particles are separated depending on the size by the electric field of the first pair of microelectrodes.
  • FIG. 2 shows the conditions during sedimentation or centrifugation. Due to the precisely adjustable centrifugal forces via the rotation speed, the particles move into the lower part of the microsystem. According to the usual centrifugation principles, the particles with the highest density sediment first. Since the particles 21 are shifted to the right by the electrical field barrier in the channel, while the particles 22 remain unaffected thereby, a separation of the two types of particles results in the channel ends 29a, 29b. The particles in each of the channel ends also arrange themselves according to their density, as in conventional centrifugation.
  • the microsystem shown can be regarded as the basic form of a device according to the invention, which basic form can be enlarged, expanded or combined with other microstructures depending on the application.
  • a microsystem according to the invention can be expanded as desired, as is known per se from dielectrophoretic microsystems. Accordingly, the channel structure can in particular have a plurality of individual channels connected to one another via branches.
  • the channels can be straight or curved. Curved channel shapes (e.g. arches, meanders, bends, angles, etc.) can be used in particular to investigate differences in the binding of particles to the channel walls.
  • the microsystem can be rotatably attached to the receptacle 12 (see FIG. 1).
  • a first centrifugation process in a first microsystem orientation, e.g. 2.
  • the orientation of the microsystem is changed by 180 °, so that the gravitational and / or centrifugal forces act counter to the arrow 23.
  • the channel ends 29a, 29b then take over the function of input depots, from which, in the presence of suitable channel structures (additional lateral branches), a further distribution of the separated particles, subgroups or a specific treatment (loading with substances, electroporation and the like) can take place.
  • orientation changes other than the 180 ° reversal mentioned are also possible.
  • the possibility of designing the receptacle 12 in such a way that the microsystem is rotated during the centrifugation.
  • FIG. 3 Another embodiment of the invention, namely a programmable loading microsystem for cells or particles, is shown in FIG. 3.
  • the centrifugation channel is divided into three parts 31a, 31b, 31c. Openings 32 are located in the intermediate walls, through which electrodes 33 on the top and bottom of the channel extend again. The openings are adapted to the particle size (typically 5 to 20 times larger than the diameter).
  • different solutions are used in each of the channel parts 31a to 31c, which serve to chemically change or load the particles.
  • the particles are inserted into a channel part (here eg 31c).
  • the particles (for example first the black, then the light ones) reach the electrodes 33 through the centrifugation and can thus be automatically transferred via the electrical field barriers through the openings 32 into the neighboring solutions.
  • microsystems can have openings (inflows, flows, outflows) that can be closed so that the particles can be easily removed or inserted after centrifugation or before.
  • all the microelectrode elements holding electrodes for particles, microfield cages, etc.
  • the method according to the invention is an electrically controlled or active centrifugation.
  • combinations with the action of optical forces (laser tweezers), magnetic forces (action on magnetic particles) or mechanical forces in the form of ultrasound forces can be provided.
  • the invention is not restricted to specific solution or suspension liquids. It is advantageous if the viscosity of the liquid contained in the microsystem is known. If the viscosity is known, the speed of rotation for setting a specific particle speed can be determined on the basis of table values or by means of a program algorithm. Alternatively, however, it is also possible to record the actual speed of the particles in the microsystem during centrifugation (e.g. with an optical sensor) and to regulate the speed in order to set a certain particle speed. It can be provided that in different sections of the channel structures, e.g. Liquids with different viscosities are contained in parallel channels that are only connected to one another via an opening. In this case, however, viscosities are preferred in which it is ensured that the diffusion of the liquids through the opening over the centrifugation period is relatively small or negligibly small.
  • the invention can be implemented in a modified manner, in that particles are introduced on the side of the microsystem facing away from the axis of rotation and under the effect of the buoyancy or under the combined effect of the buoyancy and the centrifugal forces Walk end of the microsystem.
  • the microsystem is adapted depending on the application with regard to the channel structure and the alignment of the electrode devices.
  • the cross-channel dimensions are generally much larger than the diameter of the individual particles. This advantageously avoids clogging of the channels. If only particles with particularly small dimensions are to be manipulated (eg bacteria or viruses or cell organelles), the channel dimensions can be reduced accordingly, for example to amounts below 10 ⁇ m.
  • the invention is implemented with a microsystem that is closed on at least one side.
  • the closed end can be a closed channel end, a closed collecting zone or a closed cavity in the microsystem.
  • the invention provides for the speed of the centrifuge to be increased briefly in order to detach the particles adhering and move them further.

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Dispersion Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Fluid Mechanics (AREA)
  • Physics & Mathematics (AREA)
  • Engineering & Computer Science (AREA)
  • Microelectronics & Electronic Packaging (AREA)
  • Centrifugal Separators (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Electrostatic Separation (AREA)

Abstract

Pour manipuler les particules dans un microsystème fluidique (15) dans lequel les particules contenues dans un liquide de suspension se déplacent dans un sens de référence prédéterminé, le microsystème (15) est fermé au moins à son extrémité (17a, 17b) qui se situe dans le sens de référence. Les particules se déplacent sous l'action de la force centrifuge et/ou par gravité dans le liquide de suspension au repos par rapport au microsystème (15). Les forces centrifuge et/ou de gravité sont sensiblement parallèles au sens de référence. Dans le microsystème (15), les particules sont également soumises à des forces de déviation dont la direction s'écarte de la direction de référence.
EP99931204A 1998-06-29 1999-06-28 Procede et dispositif de manipulation de particules dans un microsysteme Withdrawn EP1092144A1 (fr)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
DE19828919 1998-06-29
DE19828919 1998-06-29
DE19853658 1998-11-20
DE1998153658 DE19853658A1 (de) 1998-11-20 1998-11-20 Verfahren und Vorrichtung zur Manipulation von Partikeln in Mikrosystemen
PCT/EP1999/004468 WO2000000816A1 (fr) 1998-06-29 1999-06-28 Procede et dispositif de manipulation de particules dans un microsysteme

Publications (1)

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