JP2009097902A - Reaction control device and reaction control method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a reaction control device capable of performing highly accurately temperature control, when generating a prescribed reaction. <P>SOLUTION: A reaction processing device is equipped at least with a reaction domain A, and a heating part for heating the reaction domain A. In the reaction control device 1, the heating part is equipped at least with a light source 11 as a heating source; a photodetection part 14 for receiving heating light L2 irradiated onto the reaction domain A, and detecting at least some of an irradiation position, an area and intensity as a detection result; and an irradiation control means for controlling light irradiation from the light source based on the detection result. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a reaction control device and a reaction control method. More specifically, the present invention relates to a technique for performing heating by light irradiation when performing a predetermined reaction, and performing the heating irradiation with high accuracy.

  In recent years, a technique for performing a chemical reaction, a biochemical reaction, or the like in a fine channel has been developed. Examples of such devices include those developed as bio / chemical applications such as μTAS (Total Analysis System) and Lab-on-a-chip. If a predetermined reaction performed with a very small amount of sample can be controlled with high accuracy, the amount of the sample can be reduced, and exhaustive analysis is possible.

  For example, the PCR (Polymerase Chain Reaction) method is expected to be applied to a technique for amplifying a target DNA fragment several hundred thousand times. That is, even if a target portion of DNA that exists only in a trace amount can be amplified, a great effect can be expected in the analysis of the base sequence.

  On the other hand, in order to carry out the reaction on a micro scale, the influence of the channel surface, its surface area, etc. is large because of the scale. Therefore, in order to perform a predetermined reaction or the like on a microscale, it is required to perform highly accurate control regarding these factors and the like. Similarly, control of reaction temperature etc. needs to be highly accurate. In this regard, Patent Document 1 discloses a technique related to measurement and control of the temperature of the liquid phase in the microchip microchannel. Among them, a technique for heating a liquid phase using an infrared laser is also disclosed.

  In addition, attempts have been made to improve reaction control and reaction efficiency by measuring high functionality of the microchannel. For example, in Patent Document 2, regarding the surface treatment of a microreactor, sample introduction is facilitated by performing treatment so that the inside of the microreactor is hydrophilic and the peripheral part is water repellent. Techniques to do this are disclosed.

International Publication No. 2002/090912 Pamphlet. Japanese Patent Application Laid-Open No. 2004-333404.

  As described above, it is desired to control the predetermined reaction efficiently. In particular, it is demanded that temperature control can be performed with high accuracy. Therefore, the main object of the present invention is to provide a reaction control device capable of performing temperature control with high accuracy when performing a predetermined reaction.

First, the present invention is a reaction processing apparatus including at least a reaction region and a heating unit that heats the reaction region, and the heating unit is a light source as a heating source, and for heating the reaction region. A reaction control device including at least a light detection unit that receives light of the light and detects information of the received light as a detection result, and an irradiation control unit that controls light irradiation from the light source based on the detection result. provide. The reaction area can be directly heated by heating the reaction area by light irradiation, and the detection result can be reflected in the irradiation control means by detecting the light used for the heating by the light detection unit. it can. Thereby, accurate heating irradiation becomes possible, and more accurate temperature control becomes possible.
Next, the present invention provides a reaction control device in which the light source can irradiate light having a plurality of wavelengths, and the wavelength of the light to be irradiated can be switched by time division. Heating irradiation to the reaction region and other light irradiation can be performed in a time-sharing manner.
Subsequently, the present invention provides a reaction control device in which the irradiation control unit guides light to be detected by the detection unit to a predetermined irradiation spot in the light detection unit by a hologram. By using the hologram to guide the wavelength light to be detected to a predetermined irradiation spot, it is possible to detect light with higher accuracy.
And this invention provides the reaction control apparatus which detects the light of a some wavelength by the said photon detection part being provided with a some photodetection element at least. Thereby, it is possible to detect light having a wide range of wavelengths.
Furthermore, the present invention provides a reaction control device in which the light detection unit includes a filter that transmits light of a specific wavelength. Thereby, the light receiving accuracy can be further improved.
Further, the present invention provides the reaction control device, wherein the filter includes a plurality of regions having different specific wavelengths that can be transmitted, and the light detection unit includes a light detection element corresponding to each wavelength of the light transmitted through the filter. provide. Thereby, light of a plurality of wavelengths can be detected with high accuracy.
The present invention provides a reaction processing apparatus in which a plurality of the reaction regions are provided in an array, and the light source and the light detection unit are also provided in an array corresponding to the reaction regions. Thereby, each predetermined reaction performed in a some reaction area | region can be controlled with high precision.
Further, when performing a predetermined reaction in the reaction region, the reaction control method performs heating by irradiating at least the reaction region with light, irradiating the reaction region with heating light, and irradiating the irradiated A reaction control method is provided that at least carries out the control of the light irradiation based on the information by detecting the information of the received light, detecting the information of the received light.

  According to the present invention, temperature control can be performed with high accuracy when performing a predetermined reaction. As a result, the predetermined reaction can be controlled with high accuracy.

  Hereinafter, a reaction control device and a reaction control method according to the present invention will be described with reference to the accompanying drawings. Each embodiment shown in the accompanying drawings shows a representative example according to the present invention, and the scope of the present invention is not interpreted narrowly.

  FIG. 1 is a simplified side view of a first embodiment of a reaction control device according to the present invention. FIG. 2 is a simplified side view for explaining one state of the embodiment. The code | symbol 1 in FIG. 1 has shown the reaction control apparatus which concerns on this invention. The size and layer structure of the reaction control device 1 can be appropriately selected according to the purpose, and the configuration of the reaction control device 1 can be designed or changed within the scope of the object of the present invention. In the drawings used below, for the convenience of explanation, the configuration of the apparatus is shown in a simplified manner.

  The reaction control device 1 includes a substrate 10, a light source 11, condenser lenses 12 and 13, and a light detection unit 14. A predetermined reaction is performed in the reaction region A of the substrate 10. The reaction control device 1 can heat the sample in the reaction region A by irradiating the reaction region A of the substrate 10 with light from the heating light source 11. And this light is detected by the light detection part 14, and the light irradiation of the light source 11 is controlled based on the detection result. Thus, the light detection part 14 which detects the light irradiated with heating is provided, and irradiation control of heating irradiation can be performed based on this detection result.

  The substrate 10 is formed of a substrate body 101 and a coating layer 102, and the reaction region A is formed by laminating the substrate body 101 and the coating layer 102. The substrate body 101 has a shape conforming to the reaction region A, and a space corresponding to the reaction region A is formed by laminating a covering layer 102 thereon.

  The material of the substrate body 101 is not limited. For example, various synthetic resins and glass can be used. For example, polymethyl methacrylate (PMMA), polydimethylsiloxane (PDMS), polyethylene terephthalate (PET), polycarbonate, Borosilicate glass, quartz glass, or the like can be used. Among these, it is preferable that the material has light transmittance, and specifically, it is desirable to use polymethyl methacrylate, a low fluorescent light emitting plastic material, borosilicate glass, quartz glass, or the like. Since such a material has particularly excellent light transmittance, it is desirable in that high photothermal efficiency can be obtained.

  In the present invention, the coating layer 102 is not necessarily limited to a layered form, and is not limited to coating the substrate 10 with the coating layer 102, but is preferable because the substrate body 101 can be sealed with the coating layer 102.

  The material of the coating layer 102 is not limited. For example, various synthetic resins and glass can be used. For example, polymethyl methacrylate (PMMA), polydimethylsiloxane (PDMS), polyethylene terephthalate (PET), polycarbonate, Borosilicate glass, quartz glass, or the like can be used. Further, like the substrate body 101, it is desirable that the material has light transmittance, and specifically, it is desirable to use polymethyl methacrylate, a low fluorescent light emitting plastic material, borosilicate, quartz glass, or the like.

  Although the light source 11 is not limited, it is desirable that the light L1 to be irradiated has directivity and convergence, and preferably a semiconductor laser (LD) or a light emitting diode (LED) light source. Further, it is desirable to further provide a switching element that controls the presence or absence of light emission. Thereby, more accurate control of light emission (that is, control of whether or not heating is performed) becomes possible. Although it does not specifically limit as a switching element, For example, TFT etc. can be used. By using a TFT or the like, it is possible to greatly contribute to the integrated production with the light detection unit 14. Of course, the form of the control system performed in the present invention is not limited to a TFT or the like, and for example, a relay switch or a digital circuit may be used.

  Examples of the LD light source include those using a semiconductor element as a light source, which is also preferable because the light emission source is small. For example, a relatively long wavelength LD light source (for example, λ = 1 to 6 μm) using a semiconductor containing Ga (gallium), In (indium), As (arsenic), P (phosphorus), etc. has a water absorption efficiency. Well, it is suitable for directly heating a liquid sample or the like. Even when a short wavelength LD light source with low water absorption efficiency is used, a heat storage layer (for example, a metal film or a resin layer) that absorbs light from the LD light source is applied to the reaction region A. The same effect can be obtained.

  And an LED light source can also be used suitably similarly to LD light source. Examples of the LED light source include a diode that emits light when a current flows through the joint, and a suitable material can be appropriately selected as the light source according to a desired wavelength or the like.

  In addition, when performing optical analysis (for example, fluorescence analysis, scattered light analysis, infrared spectroscopy, ultraviolet spectroscopy, etc.) of the sample in the reaction region A, light irradiation may be performed separately in addition to the heating light. . By using one or a plurality of light sources, light having a plurality of wavelengths can be irradiated. And you may comprise a reaction control apparatus so that these lights may become a coaxial optical path. This will be described later.

  The condenser lens 12 can focus the light emitted from the light source 11. Although not shown, a diffractive optical element, a collimating lens, and the like can be provided on the optical path as necessary. By these, the lights L1 and L2 can be focused or collimated and guided.

  The numerical aperture (NA) and the like of the condenser lens 12 can be appropriately determined in consideration of the apparatus configuration, but it is preferable that the size of the reaction area A and the light irradiation spot are approximately the same. For example, if the optical spot is equivalent to the air ring diameter, the desired NA can be calculated by the relational expression of d (radius) = 0.61 × (wavelength) × NA. In addition, if the NA is large, the beam spot can be reduced but the depth of focus is shallow. If the NA is small, the depth of focus can be deep (large), but the beam spot tends to be large.

  The light irradiated to the reaction area A is collected by the condenser lens 13 and received by the light detection unit 14 and detected. The light detection unit 14 can receive the light L2 and detect information on the light. This information includes, for example, the irradiation position, irradiation area (or spot diameter), irradiation intensity, spectrum distribution, refractive index change, etc., and may be at least one of these. More preferably, it is desirable to detect a plurality of these pieces of information. Thereby, detection accuracy can be further improved. Furthermore, more accurate heating control can be performed by feeding back this information to the irradiation control means described later. The information detected here can be determined as appropriate based on the purpose of use and the configuration of the reaction control device 1, the reaction performed in the reaction region A, and the like.

  The light detection unit 14 can use a photodiode or the like as a light detection element. For example, it can be used as a CCD or MOS. As a detection element such as a laser, a multi-divided PIN photodiode such as four or six divisions, a photo IC combining a photodiode and an integrated circuit, or the like can be used. These will be described later.

  If necessary, it is desirable to provide a filter that transmits light of a predetermined wavelength between at least the light source 11 and the light detection unit 14. Thereby, since wavelengths other than the light L1 which is a detection target can be cut, detection accuracy can be further improved. It is also preferable to pass through a dichroic mirror because the influence of scattered light and the like can be eliminated. Further, by using a plurality of filters as band-pass filters that detect only an arbitrary frequency, it is possible to perform spectroscopic analysis by detecting the light frequency and light intensity with high accuracy.

  The irradiation control means is not limited as long as it can irradiate the predetermined irradiation target spot with the light L1 emitted from the light source 11. Examples of the irradiation control means for controlling the heating irradiation include adjusting the optical path of the light irradiation by controlling the position of the optical element provided on the optical path. As such an optical element, for example, a lens, a prism, a reflecting mirror, a diffraction grating, or the like can be used. By moving the optical element and performing position control, the optical path of light irradiation can be adjusted. In the reaction control device 1, it can be performed by controlling the position of the condenser lens 12 or adjusting the irradiation direction of the light source 12. By outputting the detection result of the light detection unit 14 to the irradiation control means, the heating irradiation can be controlled with high accuracy. For example, the detection result in the light detection unit 14 can be converted into digital information by an analog-digital converter (ADC) or the like and output, and irradiation control can be performed using a CPU.

  The position control of the optical element is not limited as long as the optical element can be moved to a predetermined position. For example, it may be machine control or may be performed manually. However, the reaction control apparatus according to the present invention can be suitably used as the reaction region A as a minute region. In such a case, the movement amount of the optical component is particularly minute. From such a viewpoint, it is desirable to use an electromagnetic actuator, a piezoelectric element, a stepping motor, an ultrasonic motor, or the like.

  The moving direction of the optical element such as the condenser lens 12 is desirably movable not only in the horizontal direction (XY direction) but also in the vertical direction (Z direction, optical axis direction). Thereby, the deviation of the irradiation spot in the XY direction can be eliminated, and the size of the irradiation spot (beam spot diameter) can be adjusted.

  Next, irradiation control performed by the reaction control apparatus 1 of the present invention will be described with reference to FIG. FIG. 2 is a simplified side view for explaining an example of irradiation control performed in the reaction control device 1.

  FIG. 2 shows a state in which the substrate 10 is slightly displaced in the horizontal direction (right side with respect to the drawing). In this case, the light L1 irradiated from the light source 11 is irradiated to the reaction region A and detected as light L2. However, since the substrate 10 is displaced, the irradiation spot on the reaction area A is displaced and the incident position on the condenser lens 13 is also displaced. As a result, the irradiation spot detected by the detection unit 14 is shifted by a distance d (see the enlarged region in FIG. 2). For example, in the case of FIG. 2, the irradiation spot of the light L <b> 2 is formed at the center of the light detection unit 14 if the substrate 10 having the reaction region A is an installation position that should be originally installed. However, since the substrate 10 is displaced, an actual irradiation spot is formed at a position displaced by the distance d.

  By detecting the distance d of the position deviation of the irradiation spot by the light detection unit 14, it is possible to detect how much the irradiation of the light L1 to the reaction region A is irradiated. Taking this distance d into account, the irradiation from the light source 11 can be controlled. The detection regarding the distance d can be performed by detecting the information of the light irradiation spot by the light detection unit 14. This light detection will be described below.

  Here, the case where the reaction region A is provided on the substrate 10 and the case where the substrate 10 is shifted by the distance d has been described as an example, but the present invention is not limited thereto. That is, it is only necessary that irradiation control can be performed via the irradiation control unit based on the detection result detected by the light detection unit 14, and the reaction region A does not necessarily have to be provided on the substrate. Further, the deviation of the irradiation to the reaction region A is not limited to that caused by the positional deviation of the substrate 10. Furthermore, the actual reaction temperature can be controlled, and a temperature gradient can be intentionally formed. Since this is possible in the present invention, high-precision heating control is possible. This will become clear from the explanation given below.

  FIG. 3 is a partial conceptual diagram for explaining an example of light detection performed by the reaction control device 1 according to the present invention. In FIG. 3, portions related to the light detection of the reaction control device 1 are shown, and other configurations are omitted.

  The light detection unit 14 includes a plurality of light detection elements 141 arranged in a horizontal plane (XY direction). Specifically, the light detection elements 141a, 141b, 141c, and 141d are arranged in a matrix. Note that the enlarged region in FIG. 3 is a state in which these photodetecting elements 141 are viewed from above. It is possible to detect which photodetection element receives the irradiated spot of the received light, and also its light receiving area and light receiving intensity. Thereby, the more exact position of the irradiation spot used for heating irradiation can be detected. The information detected by the light detection unit 14 is not limited to the irradiation position, the light receiving area and the light receiving intensity, such as the position where the irradiation spot exists, but other than that, the spectral distribution and the refractive index of the received light. Of course, it may be information such as changes.

  For example, in FIG. 3, the actual irradiation spot S1 is slightly shifted to the lower right from the target irradiation spot S2. First, in the case of the target irradiation spot S2, the light detection elements 141a, 141b, 141c, and 141d have a uniform light reception area and light reception intensity. However, in the actual irradiation spot S1, the light receiving areas and light receiving intensities of the light detecting elements 141a, 141b, 141c, and 141d are not uniform. Specifically, the light detection element 141d has an irradiation spot that is too wide, and the light detection element 141a has an irradiation spot that is too narrow. From these, the actual position of the irradiation spot S1 can be detected.

Therefore, here, assuming that the light receiving areas (or light receiving intensities) of the light detection elements 141a, 141b, 141c, and 141d are PD _141a , PD _141b , PD _141c , and PD _141d , The relational expression shown in (1) can be used.

上記式（１）は、光検出部１４のＸ方向において所定目標に光照射された状態を示している。同様に、Ｙ方向の位置検出においては、下記式（２）で示される関係式を用いることができる。

The above formula (1) shows a state in which the predetermined target is irradiated with light in the X direction of the light detection unit 14. Similarly, in the position detection in the Y direction, a relational expression represented by the following formula (2) can be used.

  The above equation (2) shows a state in which the predetermined target is irradiated with light in the Y direction of the light detection unit 14. Therefore, when both of the above formulas (1) and (2) are satisfied, it can be confirmed that the target spot S2 is irradiated with light at least in the horizontal direction (XY direction).

  Further, the photodetecting element may be a so-called multi-divided one. And the more photodetection elements are provided, the more accurately the position of the irradiation spot S1 can be detected. Therefore, for example, if each of the photodetecting elements 141a, 141b, 141c, and 141d is further divided into multiple matrixes, more accurate information on the irradiation spot position can be detected.

  The case where the position of the substrate 10 in the vertical direction (Z direction) is detected will be mainly described. When the substrate 10 moves in the vertical direction, the beam spot diameter changes depending on its height, which can also be detected by the light detection unit 14. This is because the photodetection element can detect the actual irradiation spot S3 that has been received, and the photodetection area can detect the photodetection area.

  For example, the case where the substrate 10 is located slightly above the position where the substrate 10 should be (that is, from the condenser lens 12 (13) side) in FIG. 3 will be described. In this case, refraction and interference occur through the optical element of the substrate 10 and the surface of the reaction region. Thereby, the incident angle to the light detection unit 14 and the like are different. For this reason, the light receiving position, the light receiving area, the light receiving intensity, and the like of the light detection elements 141a, 141b, 141c, and 141d change. By detecting this as information on the light L2, it is possible to know how close the target irradiation spot S4 is.

  The positional deviation in the vertical direction (Z direction) can be known by detecting the beam spot diameter of the irradiation spot S3. If the actual beam diameter of the irradiation spot S3 is larger than the target beam diameter of the irradiation spot S4, the substrate 10 is shifted slightly upward (that is, from the condenser lens 12 side). Of course, it is also possible to detect similarly by knowing the irradiation area of the irradiation spot S3.

  The target irradiation spot S4 in FIG. 3 is in a state where none of the light detection elements 141a, 141b, 141c, and 141d can receive light. However, the actual irradiation spot S3 covers any region of the light detection elements 141a, 141b, 141c, and 141d. From these, it is possible to detect information on the actual irradiation spot S3.

  Therefore, in the position detection in the Z direction, the substrate 10 is moved in the vertical direction while satisfying the expressions (1) and (2), and the outputs from the respective light detection elements 141a, 141b, 141c, and 141d are output. The minimum position is detected. It can be said that this position coincides with the focal position of the irradiation light.

  FIG. 4 is a partial conceptual diagram of a second embodiment of the reaction control device according to the present invention. The following description will focus on the differences from the embodiment described above, and the description of the common parts will be omitted. In FIG. 4, portions related to substrate position detection are shown, and other components are omitted.

  The code | symbol 2 shown in FIG. 4 has shown the reaction control apparatus. The reaction control device 2 includes at least a condenser lens 21 and a light detection unit 22, and performs a predetermined reaction on the substrate 20. The light detection unit 22 includes a plurality of light detection elements 221. This reaction control device 2 is characterized in that five photodetecting elements 221a, 221b, 221c, 221d, and 221e are used as the photodetecting elements 221 (see the enlarged region in FIG. 4). In this case, a light detection element divided into five may be used.

  In the light detection unit 22, a plurality of light detection elements 221a, 221b, 221c, 221d, and 221e are arranged on the horizontal plane (XY direction). It is possible to detect which photodetection element the irradiated spot of the received light is receiving, its light receiving area, light receiving intensity, spectral distribution, and the like. Thereby, more accurate information of the irradiation spot can be detected. In particular, since the light detection element 221e is arranged at the center position, it can be accurately detected even when the beam spot diameter is small.

Here, assuming that the respective light receiving areas (or light receiving intensities) of the light detecting elements 221a, 221b, 221c, 221d, and 221e are PD _221a , PD _221b , PD _221c , PD _221d , and PD _221e , The relational expression shown by the following formula (3) can be used.

  The above formula (3) shows a state in which the predetermined target is irradiated with light in the X direction of the light detection unit 22. Similarly, in the position detection in the Y direction, a relational expression represented by the following formula (4) can be used.

  The above equation (4) shows a state in which the predetermined target is irradiated with light in the Y direction of the light detection unit 22. Therefore, when both of the above formulas (3) and (4) are satisfied, it can be confirmed that the target spot S2 is irradiated with light at least in the horizontal direction (XY direction).

  Further, the description will focus on the case of detecting the position of the substrate 20 in the vertical direction (Z direction). When the substrate 20 moves in the vertical direction, the beam spot diameter changes depending on its height (see the arrow in FIG. 4), which can also be detected by the light detection unit 22. Which light detection element receives the actual irradiation spot S3 that has been received, as well as the light reception area thereof, can be detected by each of the light detection elements 221a, 221b, 221c, 221d, and 221e.

  In this case, in the position detection in the Z direction, the substrate 20 is moved in the Z direction while satisfying the expressions (3) and (4), and the outputs from the light detection elements 221a, 221b, 221c, and 221d are output. A position where the output is minimum or the output of the light detection element 221e is maximum is detected. It can be said that this position coincides with the focal position of the irradiation light.

  FIG. 5 is a partial conceptual diagram of a third embodiment of the reaction control device according to the present invention. The following description will focus on the differences from the embodiment described above, and the description of the common parts will be omitted. In addition, FIG. 3 has shown the part relevant to the light detection of a reaction control apparatus, and has abbreviate | omitted about the other structure.

  The code | symbol 3 shown in FIG. 5 has shown the reaction control apparatus. The reaction control device 3 includes at least a condenser lens 31 and a light detection unit 32. Light for measurement (or observation) is used in addition to light for heating, and light is received using the difference in refractive index of these lights. Each light has a device configuration that can be detected by the detection unit 32. In other words, the reaction control device 3 includes a plurality of light sources having different wavelengths as an optical system, and the detection unit 32 includes light detection elements corresponding to these wavelengths.

  The detection unit 32 includes a plurality of light detection elements 321, 321, 321, 321, and 322, and is characterized in that the detectable wavelengths of these light detection elements are different. The light detection element 321 is a light detection element that can detect heating light (for example, infrared light). The light detection element 322 is a light detection element that can detect measurement light (for example, blue LD). Thus, the wavelength can be changed according to the application of light (for example, for heating or measurement).

  Further, the difference in the wavelength of light causes a difference in refractive index. That is, the refractive index characteristic in the condensing lens 31 depends on the wavelength. Therefore, since the refractive indexes of the heating light and the measurement light are different, the irradiation spot diameter formed on the detection unit 32 is also different. Here, a typical example of the refractive index characteristic depending on the wavelength is shown in Table 1.

  However, when simultaneously irradiating two or more wavelengths, it is desirable to provide a filter that transmits a predetermined wavelength as necessary. Furthermore, it can also be set as the filter provided with the some area | region from which the wavelength which can permeate | transmit differs. Providing a filter can prevent the occurrence of crosstalk. For example, when infrared light is used as heating light and blue LD is used as measurement light, if these two wavelengths are irradiated simultaneously, crosstalk due to infrared light occurs on the light detection element for blue LD detection. End up. Therefore, it is desirable to provide an infrared cut filter.

  As such a cut filter, for example, it is desirable to use a reflection type such as a dichroic mirror, an absorption type such as an infrared ND filter, a hologram type using a diffraction grating, etc., and further select characteristics as required. It is desirable to do. In addition, when the hologram type is used, only a specific wavelength (for example, infrared light for heating) can be condensed at an arbitrary position, which will be described later.

Then, the irradiation spot S _{analysis of} the measurement light (eg, blue LD) (see the hatched area in FIG. 5) is smaller than the irradiation spot S _heat of the heating light (eg, infrared light). In general, it is desirable that the light detection element 322 for measurement light is provided at the center and the light detection element 321 for light for heating is provided outside because the wavelength is shorter and the refraction angle is larger. In this case, as described above, in addition to the irradiation position, the irradiation area, the irradiation intensity, and the like for the irradiation spot, the spectral distribution and the refractive index change can be suitably used. In particular, the refractive index change can be known by detecting the defocus amount.

  Moreover, when using a some wavelength for an optical system, it is desirable to control the irradiation timing. Thereby, crosstalk etc. can be prevented effectively. For example, switching control may be performed so as to shift the irradiation time of the heating light source and the position detection light source. In the switching control, a TFT or the like can be used as a switching element, so that it can be integrated with a light detection element. And it can contribute to thickness reduction, mass production, transparency, etc. of an apparatus. Of course, in the present invention, thinning, mass production, and transparency are not necessarily required, but it is desirable in that these can be realized by using TFTs.

  FIG. 6 is a partial conceptual view of a fourth embodiment of the reaction control device according to the present invention. The following description will focus on the differences from the embodiment described above, and the description of the common parts will be omitted. In addition, FIG. 6 has shown the part relevant to the light detection of a reaction control apparatus, and has abbreviate | omitted about the other structure.

  The code | symbol 4 shown in FIG. 6 has shown the reaction control apparatus. The reaction control device 4 includes at least a condenser lens 41, a diffraction grating 42, and a light detection unit 43. Lights having different wavelengths are used as heating light and measurement light, respectively. By utilizing the difference in refractive index of these lights, the light for detection can be detected by the light detection element 431 of the light detection unit 43, and the measurement light can be detected by the light detection element 432. Thereby, the light of a plurality of wavelengths can be detected by the detection unit 43, respectively.

The reaction control device 4 is characterized in that a hologram type cut filter using a diffraction grating 42 is used as an optical filter. By using such a hologram type, only a specific wavelength (for example, heating infrared light) can be condensed at a predetermined position S _heat . For example, by arranging the diffraction grating 42 designed to generate a diffraction phenomenon only at the wavelength of infrared light, the infrared light condensing spot S _heat can be formed into a desired spot shape.

  FIG. 7 is a simplified side view of the fifth embodiment of the reaction control apparatus according to the present invention. The code | symbol 5 of FIG. 7 has shown the reaction control apparatus based on this invention. The reaction control device 5 includes a substrate 50 having a reaction region A, a light source 51, a condenser lens 52, and a light detection unit 53. A predetermined reaction is performed in the reaction region A of the substrate 50. The reaction control device 5 can heat the sample in the reaction region A by irradiating the reaction region A with light from the light source 51 for heating. The substrate 50 includes a substrate body 501 and an optical element layer 502. In the optical element layer 502, optical elements 5022 are arranged corresponding to the reaction regions A, respectively. The reaction control device 5 is characterized in that the substrate 50 is provided with an optical element 5022 corresponding to the reaction region A.

  In the present invention, it is not always necessary to integrate the optical element 5022 with the substrate 50 or to provide the reaction region A on the substrate 50. However, by adopting such a structure, the position of the installation position of the substrate 50 is not necessary. Misalignment can also be detected. That is, in addition to detecting the deviation of the irradiation spots of the lights L1 and L2, the positional deviation of the substrate 50 can also be detected. Since the optical element 5022 is integral with the substrate 50 (or the substrate body 502), the optical element 5022 is also displaced in the same manner as the substrate 50 is displaced. Therefore, the position of the substrate 50 can also be detected by detecting the change in the position and size of the irradiation spot of the light L2 corresponding to the positional deviation of the optical element 5022. This position detection can use the irradiation spot detection method described above (see, for example, FIGS. 2, 3, 4, 5, 6).

  The optical element 5022 only needs to be able to introduce light into the reaction region A, and the type thereof is not limited. For example, the light L1 can be introduced into the reaction region A using a lens, a prism, a mirror, a diffraction grating, or the like as appropriate, but it is preferable to use an optical lens. If it is an optical lens, it can be easily fixed at a position corresponding to the reaction region A. There is no limitation on the material of the optical element 5022. For example, a crystal material such as silicon or calcium fluoride, a glass material such as synthetic quartz, or the like can be used. Further, a plurality of optical elements can be used together as necessary, and these can be provided on the substrate 50.

  FIG. 8 is a simplified side view of the sixth embodiment of the reaction control apparatus according to the present invention. The code | symbol 6 in FIG. 8 has shown the reaction control apparatus which concerns on this invention. The reaction control device 6 includes a substrate 60 having a plurality of reaction regions A, a plurality of heating light sources 61, condenser lenses 62 and 63, and a light detection unit 64. One feature of the reaction control device 6 is that a plurality of reaction regions A are provided, and a plurality of light sources 61 and light detection units 64 are also provided corresponding to the reaction regions A.

  In the substrate 60, a plurality of reaction areas A are formed by the substrate body 601 and the coating layer 602. The size, shape, and the like of the reaction region A are not limited, and may be different sizes and shapes, and the arrangement interval between the reaction regions A can be appropriately set as appropriate. Thus, by providing a plurality of reaction regions A, it is possible to perform comprehensive analysis on the substrate.

  For example, when the reaction region A is heated using a heating element such as a heater, heat conduction may be caused through the substrate body 601 or the like, so that the heating efficiency may be reduced or heating unevenness may easily occur. In addition to this, when a plurality of reaction regions A are used, if a heating element or the like is used for heating each reaction region A, thermal interference occurs between adjacent reaction regions A. This phenomenon may affect the temperature control and cause a temperature shift. In this regard, according to the present invention, such a thermal interference can be prevented even with the substrate 60 having a plurality of reaction regions A. Furthermore, the installation interval of the reaction area A can be shortened, and the device can be saved.

  In addition, when a semiconductor element or the like is used as the heating element, a slight temperature difference between the target temperature of the reaction region A and the actual temperature can occur. Since semiconductor elements generally have manufacturing variations, the heating amount varies for each heater unit in the substrate. In addition, semiconductor elements generally have a property that their characteristics change with temperature. For example, a MOS transistor using single crystal silicon has negative temperature characteristics, and even when the same voltage value is applied, the flowing current decreases as the temperature increases. Therefore, even if the voltage value is the same, the heating amount varies depending on the temperature. Such a phenomenon sometimes becomes a problem in temperature control.

  On the other hand, in the present invention, since accurate heating irradiation can be performed without being affected by the above-described effects, it is possible to perform accurate heating irradiation for each of the plurality of reaction regions A. In particular, by providing the light source 61 individually for each reaction region A, individual heat control of each reaction region A becomes possible. In particular, a plurality of reaction regions A can be arranged in an array, and a plurality of light sources 61 and light detection units 64 can be arranged in an array so as to correspond to each reaction region A.

  Furthermore, in the present invention, most of the energy can be absorbed by each reaction region A when performing heat irradiation. The remaining light can be detected by the light detection unit 64. At that time, the spot size of the detector can be adjusted. Therefore, if the transmitted light (detected light L2) is small, it is condensed. The sensitivity (or temperature) can be increased, and if there is a large amount of transmitted light, the sensitivity can be lowered without condensing light. Thus, in the present invention, the amount of heating can be controlled.

  In the substrate 60, a plurality of reaction areas A are formed by the substrate body 601 and the coating layer 602. The size, shape, and the like of the reaction region A are not limited, and may be different sizes and shapes, and the arrangement interval between the reaction regions A can be appropriately set as appropriate. Thus, by providing a plurality of reaction regions A, it is possible to perform comprehensive analysis on the substrate. The substrate will be described later.

  FIG. 9 is a simplified side view of the seventh embodiment of the reaction control apparatus according to the present invention. The code | symbol 7 in FIG. 9 has shown the reaction control apparatus which concerns on this invention. The reaction control device 7 includes a substrate 70 having a plurality of reaction regions A, a plurality of heating light sources 71, condenser lenses 72 and 73, and a light detection unit 74. The reaction control device 7 is characterized in that an optical element 7022 is also provided on the substrate 70 in addition to a plurality of reaction areas A, a light source 71, and a light detection unit 74.

  FIG. 10 is a simplified perspective view of one embodiment of a substrate that can be used in the reaction control apparatus according to the present invention. The code | symbol 80 of FIG. 10 has shown the board | substrate which can be used with the reaction control apparatus based on this invention. The substrate 80 includes a sample injection port 801, a flow path 802, and a discharge port 803 for collecting or discharging a reaction product after use (for example, after completion of the reaction). In FIG. 10, the flow path structure of the substrate is illustrated, and other configurations (for example, a coating layer) in the substrate and the reaction control device are omitted.

  As a method for introducing the sample into the reaction region A of the substrate 80, a commonly used technique can be appropriately employed. The sample introduced from the inlet 801 flows along the direction of the arrow F, and is eventually collected or discarded at the outlet 803. In particular, when the flow path 802, the reaction region A, or the like is a minute space, the sample can be introduced using a capillary phenomenon or the like.

  The substrate 80 can be structured such that the reaction region A is a minute space for performing a predetermined reaction, and the sample is communicated by a minute channel 802 to guide the sample to the reaction region A. In addition, the surface of these channels can be appropriately coated to increase the hydrophilicity, and the sample can be transported efficiently. In the present invention, the reaction region A is not necessarily provided on the substrate or the like, and the reaction region can be formed on a suitable member depending on the purpose of use.

  FIG. 11 is a schematic configuration diagram of an eighth embodiment of the reaction control device according to the present invention. Hereinafter, description will be made centering on differences from the above-described embodiment, and description of common parts will be omitted.

  The code | symbol 9 shown in FIG. 11 has shown the reaction control apparatus. The reaction control device 9 includes a substrate 90, light sources 91a, 91b, 91c, and 91d, corresponding half mirrors 92a, 92b, 92c, and 92d, a condensing lens 93, and a light detection unit 94. Yes. One feature of the reaction control device 9 is that it includes, as an optical system, four types of light sources and a light detection unit 94 that can receive four types of wavelengths corresponding thereto.

  Although not shown, the substrate 90 is provided with a reaction region, and this reaction region is irradiated with heat. The light source 91a is used as a heating light source. The reaction region of the substrate 90 is heated by the light emitted from the light source 91a. For example, an infrared laser diode or the like can be used.

  The light sources 91b, 91c, and 91d can be used as light sources for purposes other than heating. That is, three different wavelengths can be used as measurement light. For example, a red laser diode (RLD), a blue laser diode (BLD), and a green laser diode (GLD) can be used. The light sources 91b, 91c, and 91d are not limited to those used for measurement, and can be used for other purposes as appropriate.

  Moreover, when using a laser diode as a light source, it can also be set as the light source unit 95 which accommodated each laser diode. The light source unit 95 includes four types of laser diodes, but is not limited to four types. That is, a plurality of laser diodes can be stored in the light source unit 95 in order to generate a plurality of lights (preferably laser beams) having different wavelengths.

  As described above, in the present invention, it is possible to irradiate light having a plurality of wavelengths. Further, in the present invention, light having a plurality of wavelengths can be irradiated in a time division manner. For example, by dividing the time of heating and measurement (or observation) light, it is possible to detect this in real time while performing optimum temperature control (heating control).

  This will be described by taking as an example a case where the gene amplification reaction is performed by PCR and analyzed in real time by fluorescence detection. In the present invention, since light of a plurality of wavelengths can be irradiated in a short time, a plurality of types of fluorescent markers corresponding to these can be excited. As a result, the state of a chemical reaction such as PCR can be accurately grasped in real time. Furthermore, the detection result can be fed back to the irradiation control means in real time. That is, in this case, the progress of the reaction and the like can be grasped individually for each reaction region A based on the measurement result of fluorescence detection, and heating of each reaction region A can be individually controlled based on this.

  Of course, when only one type of fluorescent marker is used, one wavelength of excitation light may be used. However, when a plurality of fluorescent markers are used or a marker having a plurality of excitation characteristics is used, a plurality of wavelengths are divided in time. By irradiating, the characteristics with respect to each wavelength light (for example, excitation light) can be evaluated. Further, in the present invention, by irradiating light with time division, observation with a single wavelength can be performed in a shorter time, and detection sensitivity (S / N) can be improved.

  For example, in the case of the reaction control device 9, heating light (light source 91a): 1 second, first measurement light (light source 91b): 0.1 second, second measurement light (light source 91c): 0. By repeating the irradiation cycle of 1 second, third measurement light (91d): 0.1 second, heating light (light source 91a): 1 second, etc., the reaction can be observed while controlling the temperature. Can be done in real time.

Further, the light detection unit 94 can receive and detect the four wavelengths of light. In other words, the detection unit 94 is provided with a filter film including a plurality of filters that transmit light of a specific wavelength, and the specific wavelengths transmitted by these filters are different wavelengths. . In the reaction control device 9, four filters 941, 942, 943, 944 are arranged in a matrix, and these transmit the specific wavelengths λ _a , λ _b , λ _c , λ _d , respectively. As the filter film, for example, an infrared light transmission filter and an RGB filter can be used.

  In other words, it is desirable to use a filter having a plurality of regions having different specific wavelengths that can be transmitted, and to further provide a plurality of light detection elements for detecting light of each specific wavelength transmitted by the filter. More preferably, these optical systems are coaxial optical paths. First, by using the above-described filter film or the like, it is possible to cope with a wide wavelength range. In particular, when performing fluorescence analysis or the like, it is preferable because a plurality of fluorescent labels or the like can be used at the same time and can be detected with high accuracy.

  According to the reaction control device of the present invention, it is possible to accurately perform heating irradiation to the reaction region provided on the substrate. Furthermore, other optical systems such as spectroscopic measurement can be accurately irradiated in the same manner as in the heating irradiation by performing the coaxial optical path. Furthermore, in the case of a substrate having a plurality of reaction regions, it is possible to individually control each reaction region by providing a light source corresponding to each reaction region.

  Further, by providing a heating light source (for example, LD or LED) for each reaction region and driving each of these heating light sources independently, the temperature of each reaction region can be controlled individually and with high accuracy. . Thereby, it is also possible to control by changing the set temperature for each reaction region A. Further, even when performing spectroscopic measurement, if this is performed on the coaxial optical path, high irradiation accuracy can be obtained at the same time, so that an accurate measurement result can be obtained in real time. Furthermore, it is also possible to use light having a different wavelength depending on each reaction region A. Therefore, particularly excellent effects can be exhibited particularly when a predetermined reaction is performed on a substrate having a minute reaction region or a minute flow path.

  Then, various adjustments required by the reaction control device are provided as necessary, such as array-shaped parts with positional accuracy (for example, light sources such as LDs and LEDs, lens arrays, and corresponding light detection units). It can be simplified. In addition, various configurations and parts of the apparatus described so far can be miniaturized by using a semiconductor process or the like. As a result, not only can the predetermined reaction be controlled and detected in the micro space, but also the entire device can be made smaller. In addition, by using a light detection unit for each reaction region instead of a so-called image sensor as conventionally used, it is possible to detect a reaction with high accuracy while maintaining high sensitivity.

  The reaction control apparatus of the present invention can be used as various chips and various sensors as a substrate (see, for example, FIG. 10). For example, it can be used for blood analysis chips, biopolymer detection chips, and the like. Therefore, the present invention can also be applied to bio / chemical applications such as μTAS and Lab-on-a-chip, which are analysis systems using microchannels.

  Furthermore, even if it is disposable when used as a substrate, the economic burden can be reduced. Therefore, since contamination of the sample can be prevented, higher measurement accuracy can be easily obtained.

  If the measurement sample used for the substrate can be reduced in volume and amount, a comprehensive analysis can be performed on the substrate. In addition, the analysis time can be shortened and the amount of measurement sample required can be reduced. Therefore, the technology for reducing the volume and amount of various measurement samples is expected to be applied to a wide range of fields including medical treatment, environmental science, food analysis and the like.

  The reaction control apparatus of the present invention can be used for various reactions, and also for analysis of biological genomes and proteins, drug discovery support, compound synthesis and analysis such as combinatorial chemistry, environmental monitoring test, food safety inspection, etc. Can be used. Among them, it can be suitably used as a spectroscopic measuring instrument such as a gene amplification reaction for amplifying a minute sample, a microreactor, and further a flow cytometry.

  Since flow cytometry uses not only fluorescent light emission generated from a sample but also side scattered light as a detection target, it is preferable in that a plurality of light sources as described above can be used. In addition, when performing spectroscopic measurement while passing microparticles to be measured through the microchannel, the reaction control apparatus of the present invention can be used preferably from the viewpoint of high irradiation accuracy.

  In gene amplification reactions, it is necessary to accurately amplify a small sample, but in order to control the amplification reaction, it is important to perform spectroscopic measurement and temperature control accurately. In this regard, the reaction control device of the present invention is preferable in that accurate temperature control and high-precision spectroscopic measurement can be performed. Various methods are used for the gene amplification reaction depending on the starting material and the type of amplification product. Here, as an example, the case where the PCR method is performed in the reaction control apparatus of the present invention will be described.

  In the PCR method, DNA and the like can be amplified several hundred thousand times by continuously performing an amplification cycle of “thermal denaturation → annealing with a primer → polymerase extension reaction”. The PCR amplification product thus obtained can be monitored in real time for quantitative analysis of the trace nucleic acid.

  However, it is necessary to accurately control the amplification cycle in the PCR method or the like. For this purpose, highly accurate temperature control is required. When the temperature control is insufficient, there is a problem that an irrelevant DNA sequence is amplified or no amplification is observed.

  In contrast, according to the reaction control apparatus of the present invention, particularly when heating is performed by light irradiation, not only absolute temperatures such as DNA dissociation temperature, annealing temperature, and extension temperature, Various parameters such as temperature gradient and extension time can be accurately controlled. Further, it is possible to perform measurement by changing these conditions by the number of reaction regions on one substrate. Even when labeling with a fluorescent dye or the like, light of different wavelengths can be used, so that a plurality of dyes can be detected. Because of this, it is possible to easily optimize the amplification conditions, and further contribute greatly to comprehensive analysis.

  Similar to the reaction control apparatus described so far, the same effect can be obtained by the reaction control method of the present invention. That is, when performing a predetermined reaction in a reaction region, the reaction control method performs heating by irradiating at least the reaction region with light, irradiating the reaction region with heating light, and At least, the information of the received light is detected, and the light irradiation is controlled based on the information. And of course, this reaction control method can be performed in the reaction control apparatus described so far.

It is a simplified side view of 1st Embodiment of the reaction control apparatus which concerns on this invention. It is a simplified side view for demonstrating one state of the 1st Embodiment. It is a partial conceptual diagram for demonstrating an example of the light detection performed in the said 1st Embodiment. It is a partial conceptual diagram of 2nd Embodiment of the reaction control apparatus which concerns on this invention. It is a partial conceptual diagram of 3rd Embodiment of the reaction control apparatus which concerns on this invention. It is a partial conceptual diagram of 4th Embodiment of the reaction control apparatus which concerns on this invention. It is a simplified side view of 5th Embodiment of the reaction control apparatus which concerns on this invention. It is a simplified side view of 6th Embodiment of the reaction control apparatus which concerns on this invention. It is a simplified side view of 7th Embodiment of the reaction control apparatus which concerns on this invention. It is a simplified perspective view of one example of a substrate which can be used for the reaction control device concerning the present invention. It is a simplified block diagram of 8th Embodiment of the reaction control apparatus which concerns on this invention.

Explanation of symbols

1, 2, 3, 4, 5, 6, 7, 9 Reaction control device 11, 51, 61, 71, 91a, 91b, 91c, 91d Light source 14, 22, 32, 43, 53, 64, 74, 94 Light Detector A Reaction area S Irradiation spot

Claims (8)

A reaction treatment apparatus comprising at least a reaction region and a heating unit for heating the reaction region;
The heating unit is
A light source as a heating source;
A light detection unit that receives the heating light applied to the reaction region and detects information of the received light as a detection result;
Irradiation control means for controlling light irradiation from the light source based on the detection result;
A reaction control device comprising at least   The reaction control apparatus according to claim 1, wherein the light source is capable of irradiating light of a plurality of wavelengths, and the wavelength of the light to be irradiated is switched by time division.   The reaction control apparatus according to claim 1, wherein the irradiation control unit guides light to be detected by the detection unit to a predetermined irradiation spot in the light detection unit by a hologram.   The reaction control apparatus according to claim 1, wherein the light detection unit includes a plurality of light detection elements to detect light having a plurality of wavelengths.   The reaction control apparatus according to claim 1, wherein the light detection unit includes a filter that transmits light of a specific wavelength. The filter includes a plurality of regions having different specific wavelengths that can be transmitted;
The reaction control device according to claim 5, wherein the light detection unit includes a light detection element corresponding to each wavelength of light transmitted through the filter. A plurality of the reaction regions are provided in an array,
The reaction processing apparatus according to claim 1, wherein a plurality of the light sources and the light detection units are also provided in an array corresponding to the reaction region. A reaction control method of performing heating by irradiating at least the reaction region with light when performing a predetermined reaction in the reaction region,
Irradiating the reaction area with heating light,
Receiving the irradiated light for heating, detecting information of the received light,
A reaction control method for at least controlling the light irradiation based on the information.

Images