JP2009069011A - Reaction control apparatus and method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a reaction control apparatus capable of controlling positioning for reaction on a substrate at high precision. <P>SOLUTION: The reaction control apparatus for performing a preset reaction in a reaction region A on a substrate comprises a substrate 10 having reaction regions A and an optical element 1022 to guide light into the reaction regions A, a light source 14 to apply light to samples in the reaction regions A, a position detector 16 to receive light applied to the reaction region A and detect a positional deviation of the substrate 10 in the reaction control apparatus 1, and a position correcting means to correct the positional deviation of the substrate 10. <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 a predetermined reaction on a substrate.

In recent years, a fine flow path is formed on a substrate such as glass or plastic, and a chemical reaction or a biochemical reaction is performed there. If a chemical reaction, medical diagnosis, or the like can be performed in such a fine flow path, it is possible to reduce the amount of samples and perform comprehensive analysis. In particular, it is expected to be applied as a bio / chemical application such as μTAS (Total _Analysis System) which is an analysis system using a micro flow channel and Lab-on-a-chip.

  Since analysis on a micro scale becomes possible, even a sample that exists only in a minute amount can be sufficiently analyzed. For example, the PCR (Polymerase Chain Reaction) method can be applied to a method of amplifying a target DNA fragment several hundred thousand times. Since the 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.

  However, in order to carry out the reaction on a micro scale, the influence of the surface area is large because of the scale. Therefore, highly accurate reaction control is required. For example, it is necessary to control the temperature condition of the reaction with higher accuracy. 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 increasing the functionality of microchannels. In the micro scale, the flow path surface and its surface area have a great influence on the reaction. For this reason, surface treatment or chemical modification of the channel surface is performed. In Patent Document 2, regarding the surface treatment of the microreactor, the inside of the microreactor is subjected to a treatment that exhibits hydrophilicity and the peripheral portion thereof exhibits water repellency, thereby facilitating the introduction of the sample and increasing the efficiency of the reaction. Technology etc. are disclosed.

  Here, a conventional example relating to a reaction control device that performs a predetermined reaction on a flow path substrate will be described with reference to FIG. The code | symbol 8 of FIG. 12 has shown the conventional reaction control apparatus. The reaction control device 8 includes a substrate 80, a heat source light source 81a, and detection light sources 81b and 81c. For example, focusing and tracking can be performed in the light irradiation of the light source 81b.

  The substrate 80 is a substrate provided with a fine flow path for performing a predetermined reaction. The heat source light source 81a emits a heating laser for heating the sample by the photothermal effect. A sample labeled with a fluorescent dye is present in the flow path, and the reaction proceeds by being heated at a predetermined temperature. The detection light sources 81b and 81c are irradiated with excitation light that excites the fluorescent dye. The light emitted from the heating light source 81 a and the detection light sources 81 b and 81 c travels on the same optical axis, and is condensed and irradiated onto the flow path on the substrate 80 by the objective lens 83. Thereby, the sample is heated and the fluorescence is detected. By observing the emitted fluorescence with an imaging device such as a CMOS (Complementary Metal Oxide Semiconductor) 85, a predetermined reaction can be analyzed in real time.

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

  In performing the reaction on such a substrate, it is necessary to control the reaction with high accuracy. In particular, the displacement of the substrate installed in the apparatus may have a great influence on the reaction efficiency and measurement accuracy. Therefore, the present invention mainly relates to a reaction control apparatus that performs a predetermined reaction on a substrate, and provides a reaction control apparatus that can perform position control of the substrate with high accuracy.

First, the present invention provides a reaction control device that performs a predetermined reaction in a reaction region provided on a substrate, the substrate including the reaction region and an optical element that introduces light into the reaction region, and the reaction A light source that irradiates the sample in the region with light, a position detector that detects the displacement of the substrate in the reaction control device by receiving the light irradiated on the reaction region, and a displacement of the substrate. There is provided a reaction control device including a position correcting means for correcting. By providing an optical element on the substrate, the position detection unit can detect a deviation between the target spot of light irradiation and the irradiation spot actually irradiated. Thereby, it is possible to detect how much the substrate is installed with a deviation from the target installation position. And since the position shift of a board | substrate can be corrected based on this detection result, exact position control of a board | substrate is attained.
Next, this invention provides the reaction control apparatus provided with the heating part which uses the said light source as a heat source which heats the said reaction area | region. By heating the reaction region by light irradiation, the liquid sample in the reaction region or the wall surface in the vicinity thereof can be directly heated, so that the heating efficiency is good. In addition, the light source used for light irradiation can perform high-speed and high-precision energy control, and can perform high-precision temperature control.
Then, this invention provides the reaction control apparatus whose said optical element is an optical lens provided in the substrate surface. By providing the optical lens on the substrate having the reaction region, the deviation of the substrate can be detected as the deviation of the irradiation spot as it is. Thereby, the position shift of the substrate can be detected more accurately. As a result, more accurate position control is possible.
In the present invention, the position detection unit is provided with a plurality of light receiving elements for one reaction region, and provides a reaction control device that detects an irradiation spot of the light by each light receiving element. By providing a plurality of light receiving elements and receiving light, these light receiving elements can detect a more accurate irradiation spot. Thereby, more accurate position information can be detected. As a result, the position shift of the substrate can be detected more accurately.
Furthermore, the present invention provides a reaction control device in which the position detection unit includes a filter that transmits light of a specific wavelength. Since the noise of the received light can be cut, the light detection accuracy can be further improved.
The present invention further includes an irradiation control unit that includes a condensing lens between the light source and the substrate, and moves the condensing lens so that light emitted from the light source is irradiated to a predetermined target spot. And a reaction control apparatus further provided. In addition to the position correction means, by adjusting the irradiation of the optical system, it is possible to accurately control the irradiation energy when light irradiation is performed on a predetermined target spot.
The present invention further includes a measurement light source that irradiates the sample in the reaction region with measurement light, and a measurement unit that detects detection light generated by irradiating the sample with the measurement light. A control device is provided. Since measurement light can also be accurately irradiated, a reaction control device with higher measurement accuracy can be obtained.
Furthermore, a reaction control method for performing a predetermined reaction in a reaction region provided on the substrate, the substrate including the reaction region and an optical element that introduces light into the reaction region is irradiated with light. There is provided a reaction control method for at least performing detection of positional deviation of the substrate and correction of positional deviation of the substrate by receiving light applied to the reaction region by a light detection unit.

  According to the present invention, when a predetermined reaction is performed on a substrate, it is possible to control the position of the substrate 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. In addition, each embodiment shown by the attached drawing shows the typical example of this invention, and, thereby, the range of this 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. The code | symbol 1 in FIG. 1 has shown the reaction control apparatus which concerns on this invention. The size, apparatus configuration, and the like of the reaction control device 1 can be appropriately designed or changed according to the purpose of use 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 12, a condenser lens 12, and a light detection unit 16. A predetermined reaction is performed in the reaction region A of the substrate 10. The reaction control device 1 irradiates the reaction region A of the substrate 10 with the light L11 from the light source 12, and the light detection unit 16 receives the light L12 that has passed through the reaction region A. The position shift of the substrate 10 is detected by detecting how much the position where the light L12 is actually received is shifted from the position (target position) where light L12 should be received.

  The substrate 10 is formed of a substrate body 100 and an optical element layer 102 and includes a plurality of reaction regions A. The substrate body 100 has a shape corresponding to the reaction region A, and a space corresponding to the reaction region A is formed by covering the substrate body 100 with the optical element layer 102. The optical element layer 102 includes an optical element 1022.

  The material of the substrate body 100 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 them, a material excellent in light transmittance is preferable, and specifically, polymethyl methacrylate, a low fluorescent light emitting plastic material, borosilicate glass, quartz glass, and the like are preferably used.

  The optical element layer 102 includes an optical element 1022. In the present invention, any optical element may be provided on the substrate 10, and the configuration thereof is not limited. Further, the member having the optical element 1022 is not necessarily limited to the layer shape, but it is preferable because the substrate body 100 can be sealed by the optical element layer 102. Of course, the optical element 1022 may be provided directly on the substrate 10 (or the substrate body 100).

  The material of the optical element layer 102 is not particularly limited, and various synthetic resins, glass, and the like can be used, for example, polymethyl methacrylate (PMMA), polydimethylsiloxane (PDMS), polyethylene terephthalate (PET). ), Polycarbonate, borosilicate glass, quartz glass and the like can be used. Among them, a material excellent in light transmittance is preferable, and specifically, polymethyl methacrylate (PMMA), a low fluorescent light emitting plastic material, borosilicate glass, quartz glass, and the like are preferable. Is desirable.

  In the present invention, by providing the optical element 1022 on the substrate 10, it is possible to detect the positional deviation of the substrate 10 by receiving the light L <b> 12 by the light detection unit 16. That is, since the optical element 1022 is integral with the substrate 10, the optical element 1022 is displaced as well as the substrate 10 is displaced. Accordingly, the position and size of the irradiation spot of the light L12 change corresponding to the positional deviation of the optical element 1022. By detecting this, the position of the substrate 10 is detected. Therefore, it is desirable to improve the integrity by fixing the optical element 1022 to the substrate 10 (substrate body 100). Thereby, the detection accuracy of misalignment can be improved.

  The optical element 1022 only needs to be able to introduce light into the reaction region A, and the type thereof is not limited. For example, the light L11 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. It is desirable to use an optical lens as the optical element 1022 because the optical lens can be easily fixed at a position corresponding to each reaction region A. The material of the optical element 1022 is not limited, and for example, a crystal material such as silicon or calcium fluoride, a glass material such as synthetic quartz, or the like can be used. Furthermore, a plurality of optical elements can be used in combination as necessary.

  The shape, size, and the like of the substrate 10 are not limited. For example, the reaction regions A can be arranged in a matrix in the substrate 10 and the optical elements 1022 can be provided correspondingly. The substrate 10 may be an arrayed microchip. Further, the substrate 10 does not need to include a plurality of reaction regions A, and may have a configuration having only one reaction region depending on the purpose of use.

  It is desirable that the reaction area A be a micro space that is arranged in an array. By adopting such a shape, for example, the number of reaction regions A comparable to the number of human genes can be provided on the substrate 10. For example, when the reaction region A is 300 μm × 300 μm × 300 μm (27 nL capacity) and about 40,000 wells are arranged, a small device having an area of about 6 cm square can be obtained. As a result, various desired reactions and analyzes can be performed comprehensively.

  The light source 12 is not particularly limited, but it is desirable that the directivity and convergence of the light L11 to be irradiated be good, and a semiconductor laser (LD) or light emitting diode (LED) light source is preferable. In particular, when an LD light source or an LED light source is used, high-speed and high-precision energy control is possible, and more accurate temperature control is possible. And the light source 12 can select a suitable light source suitably in consideration of a use purpose etc. Therefore, the light source 12 is not limited to the purpose only for detecting the position of the substrate 10, and is more preferably used as a heat source for heating the reaction region A. Moreover, when performing optical analysis (for example, fluorescence analysis, scattered light analysis, infrared spectroscopy, ultraviolet spectroscopy, etc.) of the sample in the reaction region A, it can also be used as a measurement light source.

  In the present invention, it is particularly desirable to heat the reaction region A with the light L11 of the light source 12. Alternatively, the light source 12 may be used as a light source for position detection or measurement, and a light source for a heat source may be provided separately. For example, although the reaction region A can be heated using a heating element such as a heater, heat conduction may be reduced through the substrate body 100 or the like, which may reduce the heating efficiency. There is also a problem that adjacent reaction regions A interfere with each other due to heat conduction and are affected by temperature. Furthermore, when strict temperature control is desired, it is necessary to warm the substrate 10 to a predetermined temperature in advance.

  In this regard, in the present invention, the light heat can be directly transmitted to the sample existing in the reaction region A by heating using the photothermal effect of the light L11 emitted from the light source 12. Furthermore, according to the reaction control apparatus of the present invention, the optical system of the substrate can be controlled with higher accuracy, so that the desired irradiation spot can be accurately irradiated with light. As a result, the reaction regions A adjacent to each other do not interfere with each other, and more accurate temperature control is possible.

  In the present invention, the light L11 from the single light source 12 can be divided by a beam splitter or the like to introduce the light L11 into a plurality of reaction regions A. However, when the light source 12 is used as a heat source, the light source 12 is used. Is preferably provided separately for the reaction region A. By providing a light source for each reaction area A, each reaction area A can be heated individually. As a result, the temperature of each reaction region A can be individually controlled, so that a minute temperature shift between the reaction regions A can be corrected, or each reaction region A can be set to a different target temperature. For example, when the reaction areas A on the substrate and the optical elements 1022 corresponding to the reaction areas A are arranged in an array, the light source 12 and the light detection unit 16 corresponding to each reaction area A are also arranged in an array. It can be.

  Further, a minute temperature shift between the reaction regions A can be said to be one of the important problems when a semiconductor element or the like is used as a heat source. Semiconductor devices generally have manufacturing variations. For this reason, even when the same temperature control is performed in each reaction region A, the heating amount varies for each heater unit in the substrate 10. 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 reaction control apparatus 1, if the light source 12 is a heat source, such a problem can be solved because the irradiation efficiency is highly accurate.

  In particular, when heating control is performed by the light source 12, it is desirable to separately provide a switching element for controlling 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 (thin film transistor) 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. Of course, the form of the control system that can be performed in the present invention is not limited to the TFT, and may be, for example, a relay switch or a digital circuit.

  The LD light source is a light source that uses a semiconductor element as a light source, and is suitable from the viewpoint that the light emission source is small. For example, a relatively long wavelength LD light source (for example, λ = 1 to 3 μm) using a semiconductor containing Ga (gallium), In (indium), As (arsenic), P (phosphorus), etc. has a water absorption efficiency. Good, suitable for heating a liquid sample directly. Further, even when a short wavelength LD light source with low water absorption efficiency is used, a similar heat treatment 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. Can have an effect.

  Similarly to the LD light source, an LED light source can also be suitably used. The LED light source only needs to use a diode that emits light when a current flows through the joint as a light source, and a suitable material can be appropriately selected as the light source according to a desired wavelength.

  The condenser lens 14 can focus the light L <b> 11 emitted from the light source 12. 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 light L12 and the like can be focused or collimated to be guided.

  The numerical aperture (NA) of the condensing lens 14 can be appropriately determined in consideration of the apparatus configuration, but it is preferable that the size of the reaction region and the light irradiation spot are approximately the same. For example, if the optical spot is equivalent to the Airy ring diameter, the desired NA can be calculated by the relational expression of d (radius) = 0.61 × λ (wavelength) × NA. In addition, if the NA is large, the beam spot diameter can be reduced, but the depth of focus becomes shallow. If the NA is small, the depth of focus increases, but the beam spot tends to become large.

  The light detection unit 16 receives the light L12 from the reaction region A. The light detection unit 16 has at least one purpose of detecting the position of the substrate 10. That is, the light detection unit 16 can function as a position detection unit for the substrate 10.

  In the light detection unit 16, a photodiode or the like can be used as a light receiving element. For example, it can be used as a CCD or CMOS. As a detection element such as a laser, a multi-segment PIN photodiode of four or six divisions, a photo IC in which a photodiode and an integrated circuit are combined, or the like can be used.

  In addition, it is desirable to provide a filter that transmits light of a predetermined wavelength between the substrate 10 and the light detection unit 16 as necessary. Thereby, since wavelengths other than the light L12 which is a detection target can be cut, detection accuracy can be further improved. For example, passing through a dichroic mirror is also preferable in that the influence of scattered light or the like can be eliminated. Moreover, the optical filter can be a band pass filter that detects only an arbitrary frequency, and by using a plurality of these filters, spectral analysis can be performed by detecting the light frequency and light intensity.

  Next, the position control of the substrate 10 performed in 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 the position control of the substrate in the reaction control apparatus 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 L11 emitted from the light source 12 is applied to the reaction region A, and this is detected as the light L12. However, since the substrate 10 is displaced, the position where the light L11 is incident on the optical element 1022 is displaced. For this reason, the irradiation spot of the light L12 emitted from the reaction region A is also shifted by a distance d in the horizontal direction (see the enlarged region in FIG. 2). That is, in the case of FIG. 2, the irradiation spot of the light L <b> 12 is formed at the center of the light detection unit 16 if the substrate 10 is to 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 16, it is possible to detect how much the substrate 10 is displaced. Based on this detection result, the positional deviation of the substrate 10 can be corrected by the position correcting means. That is, the light detection unit 16 outputs the obtained detection result as position information, and corrects the installation position of the substrate so that the positional deviation of the substrate can be corrected according to the output from the light detection unit 16. It can be a position correction means. For example, the detection result of the light detection unit 16 may be converted into digital information by an analog-digital converter (ADC) or the like and output to the position correction unit, and this may be linked to the position correction unit using a CPU or the like.

  Note that the position correcting means performed in the present invention is not limited as long as it can move the substrate 10 to a predetermined position. Therefore, it may be by machine or manually. The amount of movement of the optical component in the present invention is considered to be a minute amount, but in consideration of such a case, it is possible to use a piezoelectric element, a stepping motor, an ultrasonic motor or the like that is small and excellent in controllability. desirable. This is desirable because the substrate 10 can be accurately moved to a target position and stopped. That is, it is preferable in that it has excellent positioning accuracy and has little lost motion.

  The moving direction of the substrate 10 is desirably movable not only in the horizontal direction (XY direction) but also in the vertical direction (Z direction). By moving the substrate 10 in the horizontal direction, the deviation of the irradiation spot in the XY direction can be eliminated. Further, the size of the irradiation spot (beam spot diameter) can be adjusted by moving the substrate 10 in the vertical direction.

  Furthermore, it is desirable to separately provide an irradiation control means for moving the condenser lens 16 so that the light L11 irradiated from the light source 12 is irradiated to a predetermined target spot on the substrate (reaction area A). When the light source 12 is used for detecting the position of the substrate 10, it is desirable to fix the condenser lens 16. However, when the reaction region A is heated with the light L 11, or the light L 11 is used as measurement light for spectroscopic analysis of the sample In some cases, it is desired to irradiate the target position more accurately. In such a case, light irradiation can be performed more accurately by the irradiation control means.

  The irradiation control means only needs to be able to move the condensing lens 16, and the method is not limited, but includes a so-called autofocus function or the like. For example, an electromagnetic actuator, a piezoelectric element, a stepping motor, an ultrasonic motor, or the like can be used.

  FIG. 3 is a simplified perspective view of one embodiment of a substrate used in the reaction control apparatus according to the present invention. Reference numeral 20 in FIG. 3 indicates a substrate used in the reaction control apparatus according to the present invention. The substrate 20 includes a sample inlet 202, a flow path 204, and a discharge port 206 for collecting or discharging a reactant after use (for example, after completion of the reaction). In FIG. 3, the flow path structure of the substrate is illustrated, and other configurations (for example, optical elements) and the reaction control device in the substrate are omitted.

  Of course, the positional deviation that occurs when or after the substrate 20 is set in the reaction control device (not shown) can be appropriately corrected as described with reference to FIG.

  As a method for introducing the sample into the reaction region A of the substrate 20, a commonly used technique can be appropriately employed. The sample introduced from the inlet 202 flows along the direction of arrow F, and is eventually collected or discarded at the outlet 206. In particular, when the flow path 204, the reaction region A, or the like is a minute space, a sample can be introduced using a capillary phenomenon or the like.

  The substrate 20 can have a structure in which the reaction region A undergoes a predetermined reaction to form a minute space and communicates with the minute flow path 204 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.

  FIG. 4 is a schematic configuration diagram of a second embodiment of the reaction control device according to the present invention. Hereinafter, the difference from the first embodiment will be mainly described, and the description of the common parts will be omitted.

  The code | symbol 2 shown in FIG. 4 has shown the reaction control apparatus. The reaction control device 2 includes light sources 21a, 21b, and 21c, corresponding half mirrors 22a, 22b, and 22c, a condensing lens 23, and a light detection unit 24. And the board | substrate 20 demonstrated previously is used. The reaction control device 2 is characterized in that excellent temperature control and measurement accuracy are possible by using a heating light source, a position detection light source, and a measurement light source as an optical system.

  The light source 21a is used as a heating light source. The reaction area A of the substrate 20 is heated by the light emitted from the light source 21a. For example, an infrared laser diode can be used for the heating.

  The light source 21b is used as a position detection light source. The light emitted from the light source 21b is used as light for detecting the displacement of the substrate 20. For example, a red laser diode (RLD) can be used. Therefore, in this reaction control device 2, light having a wavelength different from that of the heating light source (for example, infrared light) is used as position detection light.

  The light source 21c is used as a measurement light source. The light emitted from the light source 21c is used as measurement light for spectroscopic measurement of the sample in the reaction region A. For example, a blue laser diode (BLD) can be used. The light source for measurement 21c may be a light source corresponding to the wavelength used for measurement.

  Light emitted from these light sources is propagated coaxially through the half mirrors 22a, 22b, and 22c, respectively. Then, the light is irradiated to a predetermined position (such as the reaction region A) of the substrate 20 via the condenser lens 23. Among them, at least light emitted from the position detection light source 21 b is received by the light detection unit 24. Thereby, the position control of the board | substrate 20 mentioned above can be performed. And you may provide the optical detection part corresponding to the wavelength of the light irradiated from another light source separately. In addition, a dichroic mirror or the like may be used for the half mirrors 22a, 22b, and 22c.

  Moreover, when using a laser diode as a light source of light source 21a, 22b, 22c, it can also be set as the light source unit 25 which stored each laser diode. The light source unit 25 includes three types of laser diodes, but is not limited to three types. That is, the light source unit 25 can store a plurality of laser diodes in order to generate a plurality of lights (preferably laser beams) having different wavelengths.

  FIG. 5 is a partial conceptual diagram of a third embodiment of the reaction control device according to the present invention. FIG. 6 is a partial conceptual diagram for explaining the position control of the embodiment. The following description will focus on the differences from the embodiment described above, and the description of the common parts will be omitted. 5 and 6 show portions related to substrate position detection, and other configurations are omitted.

  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 36 and a light detection unit 37 as a position detection unit, and performs a predetermined reaction on the substrate 30. The reaction control device 3 is characterized in that a plurality of light receiving elements 371a, 371b, 371c, and 371d are arranged as the light detection unit 37 (see the enlarged region in FIG. 5).

  In the light detection unit 37, a plurality of light receiving elements 371a, 371b, 371c, and 371d are arranged on a horizontal plane (XY direction). That is, the enlarged regions in FIGS. 5 and 6 are in a state where the light receiving element 371 is viewed from above. It is possible to detect which light receiving element receives the irradiated spot of the received light, as well as its light receiving area and light receiving intensity. Thereby, a more accurate position of the irradiation spot can be detected.

  For example, in FIG. 5, 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 receiving elements 371a, 371b, 371c, and 371d have a uniform light receiving area and light receiving intensity. However, in the actual irradiation spot S1, the light receiving area and the light receiving intensity of the light receiving elements 371a, 371b, 371c, and 371d are not uniform. Specifically, the irradiation spot of the light receiving element 371d is too wide, and the irradiation spot of the light receiving element 371a is too narrow. From these, the actual position of the irradiation spot S1 can be detected.

Therefore, here, if the light receiving areas (or light receiving intensities) of the light receiving elements 371a, 371b, 371c, and 371d are PD _371a , PD _371b , PD _371c , and PD _371d , the following equation ( The relational expression shown in 1) can be used.

  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 37. 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 37. 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).

  The light receiving element may be a so-called multi-divided one. Then, the more light receiving elements are provided, the more accurately the position of the irradiation spot S1 can be detected. Therefore, for example, if each of the light receiving elements 371a, 371b, 371c, 371d is further divided into a matrix, a more accurate irradiation spot position can be detected.

  In FIG. 6, the case where the reaction control device 3 detects the position of the substrate 30 in the vertical direction (Z direction) will be mainly described. When the substrate 30 moves in the vertical direction, the beam spot diameter changes depending on its height (see the arrow in FIG. 6), which can also be detected by the light detection unit 37. The light receiving element detects which light receiving element receives the actual irradiation spot S3 received, and further detects the light receiving area and the like.

  For example, a case where the substrate 30 is located slightly above the position where the substrate 30 should be (ie, from the condenser lens 36 side) in FIG. 6 will be described. In this case, refraction and interference occur through the optical element of the substrate 30 and the reaction region surface. Thereby, the incident angle to the light detection unit 37 and the like are different. For this reason, the light receiving position, light receiving area, and light receiving intensity of the light receiving elements 371a, 371b, 371c, and 371d change. By detecting this, it is possible to know the irradiation spot S4.

  Accordingly, 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 30 is shifted slightly upward (that is, from the condenser lens 36 side). Of course, it is also possible to detect similarly by knowing the irradiation area of the irradiation spot S3.

  In the case of FIG. 6, regarding the target irradiation spot S4, none of the light receiving elements 371a, 371b, 371c, and 371d can receive light. However, in the actual irradiation spot S4, the irradiation spot S3 also covers the areas of the light receiving elements 371a, 371b, 371c, and 371d. From these, the actual position of the irradiation spot S1 can be detected.

  Accordingly, in the position detection in the Z direction, the substrate 30 is moved in the Z direction while satisfying the expressions (1) and (2), and the outputs from the respective light receiving elements 371a, 371b, 371c, and 371d are minimized. The position that becomes is detected. It can be said that this position coincides with the focal position of the irradiation light.

  FIG. 7 is a partial conceptual view of a fourth embodiment of the reaction control device according to the present invention. FIG. 8 is a partial conceptual diagram illustrating position detection according to the embodiment. The following description will focus on the differences from the embodiment described above, and the description of the common parts will be omitted. 7 and 8 show portions related to substrate position detection, and other components are omitted.

  The code | symbol 4 shown in FIG. 7 has shown the reaction control apparatus. The reaction control device 3 includes at least a condenser lens 46 and a light detection unit 47 as a position detection unit, and performs a predetermined reaction on the substrate 40. The reaction control device 4 is characterized in that five light receiving elements 471a, 471b, 471c, 471d, and 471e are arranged as the light detection unit 47 (see the enlarged region in FIG. 7). In this case, a light receiving element divided into five may be used.

  In the light detection unit 47, a plurality of light receiving elements 471a, 471b, 471c, 471d, 471e are arranged on a horizontal plane (XY direction). It is possible to detect which light receiving element receives the irradiated spot of the received light, its light receiving area and light receiving intensity. Thereby, a more accurate position of the irradiation spot can be detected. In particular, since the light receiving element 471e is arranged at the center position, it can be accurately detected even when the beam spot diameter is small.

Therefore, here, if the light receiving areas (or light receiving intensities) of the light receiving elements 471a, 471b, 471c, 471d, 471e are PD _471a , PD _471b , PD _471c , PD _471d , and PD _471e , 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 37. 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 37. 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).

  In FIG. 8, the case where the reaction control device 4 detects the position of the substrate 40 in the vertical direction (Z direction) will be mainly described. When the substrate 40 moves in the vertical direction, the beam spot diameter changes depending on its height (see the arrow in FIG. 8), which can also be detected by the light detection unit 47. By which light receiving element the actual irradiation spot S3 received is received, and the light receiving area thereof can be detected by the light receiving element.

  In this case, in the position detection in the Z direction, the substrate 40 is moved in the Z direction while satisfying the expressions (3) and (4), and the outputs from the respective light receiving elements 471a, 471b, 471c, 471d are output. A position at which the output is minimized or the output of the light receiving element 471e is maximized is detected. It can be said that this position coincides with the focal position of the irradiation light.

  FIG. 9 is a partial conceptual diagram of the fifth embodiment of the reaction control apparatus 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. FIG. 9 shows a portion related to the detection of the position of the substrate, and other components are omitted.

  The code | symbol 5 shown in FIG. 9 has shown the reaction control apparatus. The reaction control device 5 includes at least a condenser lens 56 and a light detection unit 57 as a position detection unit. At least the light for heating and the light for position detection are used, and the difference between the refractive indexes of these lights is used, so that the detection unit 57 can detect each light. That is, the reaction control device 5 includes a plurality of light sources having different wavelengths as an optical system, and includes a light receiving element corresponding to these wavelengths in the detection unit 57, respectively.

  The detection unit 57 includes a plurality of light receiving elements 571 and 572, and is characterized in that the wavelengths that can be detected by these light receiving elements are different. The light receiving element 571 is a light receiving element capable of detecting heating light (for example, infrared light). The light receiving element 572 is a light receiving element capable of detecting position detection light (for example, blue LD).

  A difference in the wavelength of light causes a difference in refractive index. That is, the refractive index characteristic in the condensing lens 56 depends on the wavelength. Therefore, the wavelength differs depending on the light used (for example, heating light, position detection light, measurement light, etc.). Therefore, since the refractive indexes of the heating light, the position detection light, and the measurement light are different, the irradiation spot diameter formed on the detection unit 57 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 position detection light, if these two wavelengths are simultaneously irradiated, crosstalk due to infrared light occurs on the light receiving 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. Further, when the hologram type is used, only a specific wavelength (for example, heating infrared light) can be condensed at an arbitrary position. This will be described later.

Then, the irradiation spot S _analysis (see the shaded area in FIG. 9) of the position detection light (for example, blue LD) is smaller than the irradiation spot S _heat of the heating light (for example, infrared light). In general, it is desirable to provide the light receiving element 572 for the spectral analysis light at the center and the light receiving element 571 for the heating light to the outside because the wavelength is shorter and the refraction angle is larger.

  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. For switching control, a TFT or the like can be used as a switching element, so that integration with a light receiving element is also possible. 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. 10 is a partial conceptual diagram of the sixth embodiment of the reaction control apparatus 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. FIG. 10 shows a portion related to the detection of the position of the substrate, and other components are omitted.

  The code | symbol 6 shown in FIG. 10 has shown the reaction control apparatus. The reaction control device 6 includes at least a condenser lens 66, a light detection unit 67, and a diffraction grating 68 as a position light detection unit. The apparatus has a configuration in which light for heating and light for measurement are used, and the difference in refractive index between these lights is used to detect any light by the light detection unit 67. In the light detection unit 67, the light receiving element 671 detects heating light, and the light receiving element 672 detects measurement light.

The reaction control device 6 is characterized in that a hologram type cut filter using a diffraction grating 68 is used as an optical filter. By using such a hologram type cut filter, 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 68 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. 11 is a schematic configuration diagram of a seventh 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 7 shown in FIG. 11 has shown the reaction control apparatus. The reaction control device 7 includes a substrate 70, light sources 71a, 71b, 71c, 71d, corresponding half mirrors 72a, 72b, 72c, 72d, a condensing lens 73, and a light detection unit 74. Yes. One feature of the reaction control device 7 is that it includes, as an optical system, four types of light sources and a light detection unit 74 that can receive four types of wavelengths corresponding thereto.

  The light source 71a is used as a heating light source. The reaction region of the substrate 70 is heated by the light emitted from the light source 71a. For example, an infrared laser diode or the like can be used.

  The light source 71b is used as a position detection light source. The light emitted from the light source 71b is used as light for detecting the positional deviation of the substrate 70. For example, a red laser diode (RLD) can be used.

  The light sources 71c and 71d are used as measurement light sources. That is, two different wavelengths can be used as measurement light. For example, a blue laser diode (BLD) and a green laser diode (GLD) can be used. The light sources for measurement 71c and 71d may be light sources corresponding to the wavelengths used for measurement.

Further, the light detection unit 74 can receive and detect the four wavelengths of light. In other words, the light detection unit 74 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 7, four filters 741, 742, 743, and 744 are arranged in a matrix, and these transmit the specific wavelengths λ _a , λ _b , λ _c , and λ _d , respectively. For example, an RGB filter and an infrared light transmission filter can be used as the filter film.

  Thus, a series of operations such as heating control of the reaction region, position control of the substrate 70, spectroscopic measurement of the sample, and the like can be performed on the coaxial optical path. And it can respond also to a wide wavelength range by using an above described filter film | membrane together. 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.

  The reaction control apparatus according to the present invention can accurately control the position of the substrate. Furthermore, by performing heating and measurement using an optical system, these irradiations can be performed accurately. 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.

  For example, by providing a heating light source (for example, LD or LED) for each reaction region and driving these heating light sources independently, the temperature of each reaction region can be individually controlled. Thereby, it is also possible to control by changing the set temperature for each reaction region A. Even in the case of performing spectroscopic measurement, accurate measurement results can be obtained in real time because the irradiation accuracy of the measurement light is high. Furthermore, light having a different wavelength depending on each reaction region A can be used as measurement light. 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.

  And by using an array-like component with position accuracy (for example, a light source such as an LD or LED, a lens array, and a corresponding light detection unit), various adjustments necessary for the reaction control device are simplified. Can be 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 a predetermined reaction / detection be made in the micro space, but also the entire apparatus can be made smaller. Further, 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 device of the present invention can be used in the form of various chips and various sensors as a substrate. 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, since the spectroscopic measurement is performed while passing the microparticles to be measured through the microchannel, the reaction control device of the present invention can be suitably used 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 with the reaction control method of the present invention. In a reaction control method for performing a predetermined reaction in a reaction region provided on a substrate, light is irradiated to the reaction region and a substrate including an optical element that introduces light into the reaction region. It is a method of at least performing detection of positional deviation of the substrate and correction of positional deviation of the substrate by receiving irradiated light at a light detection unit. 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 the position control of the board | substrate in the said 1st Embodiment. It is a simplified perspective view of one example of a substrate used for a reaction control device concerning the present invention. It is a schematic block 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 for demonstrating the position control of the board | substrate in the embodiment. It is a partial conceptual diagram of 4th Embodiment of the reaction control apparatus which concerns on this invention. It is a partial conceptual diagram for demonstrating the position control of the board | substrate in the embodiment. It is a partial conceptual diagram of 5th Embodiment of the reaction control apparatus which concerns on this invention. It is a partial conceptual diagram of 6th Embodiment of the reaction control apparatus which concerns on this invention. It is a schematic block diagram of 7th Embodiment of the reaction control apparatus which concerns on this invention. It is a schematic block diagram of an example of the conventional reaction control apparatus.

Explanation of symbols

1, 2, 3, 4, 5, 6, 7 Reaction control device 10, 20, 30, 40, 70 Substrate 12, 21a, 21b, 21c, 71a, 71b, 71c, 71d Light source 16, 24, 37, 47, 57, 67, 74 Photodetector 1022 Optical element A Reaction area S Irradiation spot

Claims (8)

A reaction control device for performing a predetermined reaction in a reaction region provided on a substrate,
A substrate comprising the reaction region and an optical element for introducing light into the reaction region;
A light source for irradiating the sample in the reaction region with light;
A position detection unit that detects a positional shift of the substrate in a reaction control device by receiving light irradiated to the reaction region;
Position correcting means for correcting the positional deviation of the substrate;
A reaction control device comprising:   The reaction control apparatus according to claim 1, further comprising a heating unit that uses the light source as a heat source for heating the reaction region.   The reaction control apparatus according to claim 1, wherein the optical element is an optical lens provided on a substrate surface.   The reaction control apparatus according to claim 1, wherein the position detection unit is provided with a plurality of light receiving elements for one reaction region, and detects the irradiation spot of the light by each light receiving element.   The reaction control apparatus according to claim 1, wherein the position detection unit includes a filter that transmits light of a specific wavelength.   The apparatus further comprises a condensing lens between the light source and the substrate, and further includes an irradiation control means for moving the condensing lens so that light emitted from the light source is irradiated to a predetermined target spot. The reaction control apparatus according to claim 1, characterized in that: A measurement light source for irradiating the sample in the reaction region with measurement light;
A measurement unit for detecting detection light generated by irradiating the sample with the measurement light;
The reaction control device according to claim 2, further comprising: A reaction control method for performing a predetermined reaction in a reaction region provided on a substrate,
Irradiating the substrate with the reaction region and an optical element that introduces light into the reaction region,
By detecting the light irradiated to the reaction region by a light detection unit, the positional deviation of the substrate is detected,
A reaction control method for performing at least correcting the positional deviation of the substrate.

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