JP2013000012A - Reaction device, and reactor using the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a reaction device enabling minute amounts of nucleic acid to be analyzed, eliminating waste of reaction devices in batch treatment and waiting-time, and enabling simple fluorescence detection and high-speed temperature change to be made compatible with each other.SOLUTION: A plurality of reaction devices in each of which a plurality of reaction wells are formed on a metallic thin plate of high thermal responsiveness, wherein all of the reaction wells are laid on an identical circumference and recessed parts are formed at the bottoms thereof respectively. Because of enabling minute amounts of a specimen to be held in the recessed parts of the reaction wells, the contact area of the metallic thin plate with the minute amounts of specimen increases, also resulting in higher efficiency of thermal response.

Description

  The present invention relates to a structure of a reaction device for analyzing a biological sample by amplifying a nucleic acid contained in the biological sample, and a reaction apparatus using the structure.

  Analysis of nucleic acids contained in biological samples such as blood, plasma, and tissue fragments not only in academic research such as biology, biochemistry, and medicine, but also in various fields such as industries such as diagnosis, crop varieties improvement, and food inspection It is done in The most widely used method for analyzing nucleic acids is a technique called PCR (Polymerase Chain Reaction), which specifically amplifies the nucleic acid in the region to be analyzed. As an application of PCR, it is possible to detect a very small amount of nucleic acid with high sensitivity by attaching a fluorescent label to a nucleic acid to be analyzed, irradiating excitation light and measuring the fluorescence intensity over time.

  In PCR, a cycle comprising 30 steps of heating a solution containing a nucleic acid and a reagent for amplifying it to about 95 ° C. to thermally denature the nucleic acid and then cooling to about 60 ° C. to proceed with nucleic acid annealing and extension reaction. Repeated ~ 40 times. In the current mainstream PCR apparatus, a reaction device called a microtiter plate having 96 to 384 reaction wells is arranged on a Peltier element, and a temperature cycle is given by raising and lowering the temperature of the Peltier element. In this method, since it takes time to change the temperature of the Peltier element itself, it has been a big problem for shortening the analysis time.

  Non-Patent Document 1 has a structure in which a disk-type reaction device having a reaction well rotates in contact with a heater set at a plurality of temperatures in advance in order to solve the problem of speeding up the temperature cycle. It is not necessary to change the temperature of the heater, and the temperature of the reaction device can be changed quickly. However, this structure has a problem that the entire reaction device is made of a plastic having a low thermal conductivity, and the thermal response is deteriorated when the reaction device is rotated in contact with a heater having a different set temperature.

  As a method for solving this problem, for example, as disclosed in Patent Document 1, it is conceivable to use a metal material having a high thermal conductivity for the surface in contact with the heater. In Patent Document 1, a metal material is proposed as a plastically deformable material for the main purpose of plugging the conduit, but it is also a useful structure from the viewpoint of thermal response.

  However, even in the document described in Patent Document 1, in the case of a very small amount of sample, there is a problem that the sample becomes spherical and the area in contact with the metal material is reduced, resulting in poor thermal response.

JP-T-2004-516127

Tsuguto Fujimoto, et al., Jpn.J.Ingect.Dis., 63,31-35 (2010)

  An object of the present invention is to provide a reaction device having a high thermal response even with a small amount of sample.

  As a method for solving the above-described problem, the reaction device of the present invention includes a solution storage unit that stores a solution, and a thermally conductive first layer at least on the bottom side of the solution storage unit. Having a recess for holding.

  By using the reaction device according to the present invention, the contact area between a small amount of the sample and the metal thin plate is increased by forming the recess at the bottom of the reaction well, and it is possible to increase the efficiency of the thermal response.

It is a top view which shows the reaction device to which this invention is applied. It is the schematic diagram which mounted the reaction device to which this invention was applied to the reaction apparatus. It is the schematic diagram which mounted the reaction device to which this invention was applied only partially in the reaction apparatus. It is a top view which shows the reaction device to which this invention is applied. It is a top view which shows the reaction device to which this invention is applied. It is a top view which shows the reaction device to which this invention is applied. It is a figure which shows the cross-sectional structure of the reaction device as a comparative example of this invention. It is a figure which shows the cross-sectional structure of the reaction device as a comparative example of this invention. It is a figure which shows the planar structure of the reaction device as a comparative example of this invention. It is a figure which shows the 1st cross-sectional structure of the reaction device to which this invention is applied. It is a figure which shows the planar structure of the reaction device of FIG. It is an enlarged view of the reaction device shown in FIG. It is a figure which shows the cross-sectional structure of the reaction device as a comparative example of this invention. It is sectional drawing which shows 2nd Embodiment of the reaction device to which this invention is applied. It is a perspective view of 2nd Embodiment of the reaction device to which this invention is applied. It is sectional drawing which shows 3rd Embodiment of the reaction device to which this invention is applied. It is an enlarged view of the reaction device shown in FIG.

  Hereinafter, the structure of a reaction device according to the present invention and a reaction apparatus using the same will be described in detail with reference to the drawings.

  FIG. 1 is a plan view of the reaction device of the present invention. However, the characteristic configuration of the present invention will be described in the following first to third embodiments. In the reaction device 1, a plurality of reaction wells 2 are formed on an arc having a radius r1. Although the number of reaction wells 2 is eight in FIG. 1, it may be more or less than that. The planar shape of the reaction well 2 is also a circle in FIG. 1, but may be a polygon. Here, the intervals d1 between the adjacent reaction wells 2 are all constant. The interval d1 does not necessarily have to be constant, but when the fluorescence intensity of the reaction well 2 is measured by the reaction apparatus, signal processing becomes easier when the interval d1 is constant. . The outer shape of the reaction device is an outer boundary 10 that is concentric with the center o of the arc having the radius r1 and having a radius r2 smaller than the radius r1, and a radius r3 that is concentric with the center o of the arc having the radius r1 and larger than the radius r1. The outer boundary 11 is an arc and the outer boundary 12 is an arc having a radius r4 tangent to the radius r2 and the radius r3. Here, the distance d2 between the intersection point 13 where the outer boundary 12 which is an arc of radius r4 intersects the arc of radius r1 and the reaction well 2 closest to the intersection point 13 is set to be half or less of d1. By doing in this way, it becomes possible to arrange | position the several reaction device 1 on the circumference which has the same radius as the circular arc of radius r1, without interfering.

  FIG. 2 schematically shows an example in which a plurality of reaction devices 1 shown in FIG. 1 are arranged in a reaction apparatus 5 that performs PCR by contact rotation on a heater set to a plurality of temperatures. The reaction device 1 rotates on a circumference having the same radius as the radius r1. Although the control of the rotational movement may be set freely, it is desirable to make the rotational movement constant speed so that all reaction wells 2 on the reaction device 1 can have the same temperature history. Although FIG. 2 shows an example in which eight reaction devices 1 can be arranged, the number of reaction devices that can be mounted on the maximum can be freely set depending on the design of the reaction device 1. In the case of the reaction device of the present invention, since all the reaction wells 2 are arranged on the circumference of the same radius r1, the optical system 29 that measures the fluorescence intensity by irradiating the reaction well 2 with excitation light is used. If all the reaction wells 2 pass under the optical system 29, the fluorescence intensity of all the reaction wells 2 can be measured. Although FIG. 2 shows an example in which there is one optical system 29, it may be necessary to arrange a plurality of optical systems so as to be compatible with a plurality of types of fluorescent labels. In that case, similarly to the optical system 29, it can be arranged at any other location on the circumference of the same radius r1.

  The heaters 21 to 28 arranged on the lower surface of the reaction device 1 are controlled to individual temperatures. For example, if the heaters 21 and 22 are controlled to 95 ° C. and the heaters 23 to 28 are controlled to 60 ° C., When the reaction device is moved in contact with the heater 21 and the heater 22, the nucleic acid is thermally denatured, and when the reaction device is moved in contact with the heater 23 to 28, the annealing and elongation reaction of the nucleic acid proceeds. One cycle of PCR can be performed by moving the device once in the circumferential direction. FIG. 2 shows an example in which the reaction device 1 is arranged on all the eight heaters 21 to 28. However, when the number of samples to be analyzed is small, the number of reaction devices is reduced and the required number of reactions is obtained. It is possible to perform analysis only with the device.

  FIG. 3 shows a case where the number of samples to be analyzed is three. If three reaction devices 1 are set in the reaction apparatus 5, the same analysis as in FIG. 2 is possible. In FIG. 3, the three reaction devices are all arranged adjacent to each other, but are not necessarily adjacent to each other, and the arrangement position can be freely determined. As described above, in the present invention, the number of reaction devices can be freely changed according to the number of samples to be analyzed, and the waste of reaction devices can be reduced as compared with the case of batch processing with a disk-type reaction device. Become.

  When PCR is started, it is necessary to apply the temperature cycle 30 to 40 times as described above, and during that time, the three reaction devices 1 continue to rotate on the heaters 21 to 28. However, after starting the analysis, a new sample may be generated. In the case of batch processing with a disk-type reaction device, the next sample cannot be set unless the started analysis is completed. However, in the case of the present invention, the reaction device 1 is added if there is an empty space. It is possible to insert. Similarly, if there is a sample that has been analyzed, it can be discharged from the heater at any time even if the reaction device being analyzed is rotating on the heater. That is, it is possible to perform continuous loading / unloading in which samples to be analyzed are sequentially inserted on the heater and finished samples are sequentially discharged from the heater. As described above, in the present invention, once an analysis is started, it is possible to eliminate a waiting time that the next analysis cannot be started until the analysis is completed.

  In the present invention, all reaction wells 2 move on the circumference of the same radius r 1 and measure the fluorescence intensity at the timing when they pass under the optical system 29. When the reaction device 1 is rotating at a constant speed, if the moving speed is increased, the measurement time assigned to each reaction well 2 is shortened accordingly, so that it is difficult to obtain a sufficient fluorescence intensity. There is.

  FIG. 4 is a plan view showing a reaction device different from FIG. 1 to which the present invention is applied. In order to solve the above problems, the reaction well 2 has an elliptical structure in which the shape is curved along the radius r1. By making the structure of the reaction well 2 into a curved ellipse structure as shown in FIG. 4, the time required for each reaction well to pass under the optical system 29 becomes longer even in the case of constant-speed rotation movement. It is also possible to increase the fluorescence intensity that can be produced. Also in this case, the distance d2 between the intersection point 13 where the outer boundary 12 which is an arc of radius r4 intersects the arc of radius r1 and the reaction well 2 closest to the intersection point 13 is set to be less than half of the distance d1 between the adjacent reaction wells 2. deep. By doing in this way, it becomes possible to arrange | position the several reaction device 1 on the circumference which has the same radius as the circular arc of radius r1, without interfering.

  FIG. 5 is a plan view showing a reaction device to which the present invention is applied. In the reaction device 1, a plurality of reaction wells 2 are formed on an arc having a radius r1. Although the number of reaction wells is eight in FIG. 5, it may of course be larger or smaller than that. The planar shape of the reaction well 2 is also a circle in FIG. 5, but may be a polygon. Here, the intervals d1 between the adjacent reaction wells 2 are all constant. The interval d1 does not necessarily have to be constant, but when the fluorescence intensity of the reaction well 2 is measured by the reaction apparatus, signal processing becomes easier when the interval d1 is constant. . The external shape of the reaction device is tangent to the external boundary 10 and the external boundary 11 which are straight lines parallel to each other, the external boundary 14 perpendicular to the external boundary 10 and the external boundary 11, and the external boundary 10 or the external boundary 11 and the external boundary 14. The outer boundary 12 is an arc having a radius r4. Here, the distance d2 between the intersection point 13 where the outer boundary 12 which is an arc of radius r4 intersects the arc of radius r1 and the reaction well 2 closest to the intersection point 13 is set to be half or less of d1. By doing in this way, it becomes possible to arrange | position the several reaction device 1 on the circumference which has the same radius as the circular arc of radius r1, without interfering. Thus, by making the shape of the reaction device 1 substantially rectangular, it is possible to sandwich and hold two parallel sides when handling the reaction device, and it is possible to handle the reaction device more stably.

  FIG. 6 is a plan view showing a reaction device to which the present invention is applied. The basic structure is the same as in FIG. 4 except that two reaction wells 2 that are closest to the intersection 13 where the outer boundary 12 that is an arc of radius r4 intersects the arc of radius r1 are omitted, and the hole 3 is formed instead. It is arranged near the boundary. Even when the reaction wells are omitted in this way, it is desirable to keep the interval d1 between the reaction wells constant. This is because signal processing can be facilitated if the positional information of the missing reaction well is known. The center position 15 of the reaction well closest to the intersection 13 that should have existed when the reaction wells were arranged at the interval d1 between the adjacent reaction wells 2 and the outer boundary 12 that is an arc of radius r4 are the radius r1. The interval d2 of the intersection 13 that intersects the arc is set to be less than half of d1. By doing in this way, it becomes possible to arrange | position the several reaction device 1 on the circumference which has the same radius as the circular arc of radius r1, without interfering. Although four holes 3 are formed in FIG. 6, it may be more or less than that. The planar shape of the hole 3 is also a circle in FIG. 6, but it may be a polygon. Furthermore, although the hole 3 is penetrated in FIG. 6, it is not necessarily required to penetrate. In FIG. 6, two reaction wells 2 closest to the intersection 13 where the outer boundary 12, which is an arc having a radius r 4, intersects the circular arc having the radius r 1 are missing for the convenience of arranging the hole 3 in the vicinity of the outer boundary of the reaction device 1. However, the hole 3 may be arranged at a position where the reaction well 2 does not need to be deleted, or the reaction well near the center of the reaction device 1 may be deleted and the hole 3 may be arranged near the center. Absent. If a hole is formed in a part of the reaction device, it can be fixed by, for example, inserting a pin into the hole 3 when handling the reaction device, and the reaction device 1 can be handled more reliably. It becomes.

  Next, the details of the cross-sectional structure of the reaction device 1 will be described. Prior to the description of this reaction device, a reaction device as a comparative example will be described with reference to FIG.

  FIG. 7 is a diagram showing a cross-sectional configuration of a reaction device as a comparative example of the present invention. In the reaction device 100, the thin plate portion 410, the thick plate portion 420, and the lid 430 are roughly composed of three layers in order from the lower surface side. The thick plate portion 420 is provided with a through hole, and the side wall 400 and the thin plate portion 410 constitute the reaction well 200. A sample solution 460 and oil 470 are contained in the reaction well 200. And between the thin plate part 410 and the thick plate part 420 and between the thick plate part 420 and the lid | cover 430, it has the adhesive layers 440 and 450 for bonding both together. The thin plate portion 410 has a flat shape.

  However, if a limited sample is distributed to many reaction wells for multi-item analysis, the sample solution that can be distributed to each reaction well 200 decreases as the number of items increases. When the amount of the sample solution 460 stored in the reaction well 200 becomes extremely small, the sample solution 460 cannot cover the entire bottom surface 600 of the reaction well 200 as shown in FIG. 7 (comparative example). The sample solution 460 becomes spherical as in the example) (thin adhesive layers 440 and 450 are not shown). When the bottom surface 600 has a planar shape, it is difficult to fix the spherical sample solution 460 at a desired position. As shown in FIG. 9 (comparative example), the position of the sample solution 460 is on the circumference having the radius r1. There is a concern that the fluorescence measurement cannot be performed due to the deviation from the irradiation detection region 700 of the optical system disposed in the optical system. In addition, when the sample solution 460 is spherical, a region in contact with the bottom surface 600 is reduced, which causes a problem that efficiency of heat transfer performed through the thin plate portion 410 is reduced.

[First Embodiment]
FIG. 10 is a diagram showing a first cross-sectional configuration of the reaction device 1 to which the present invention is applied. The reaction device 1 includes a thin plate portion 41, a thick plate portion 42, and a lid 43 in order of three layers in order from the lower surface side. And between the thin plate part 41 and the thick plate part 42 and between the thick plate part 42 and the lid | cover 43, it has the adhesive layers 44 and 45 for bonding both together. The adhesive layers 44 and 45 may be omitted as long as the thin plate portion 41 and the thick plate portion 42 and the thick plate portion 42 and the lid 43 can be directly bonded.

  Further, the thick plate portion 42 is provided with a through hole, and the side wall 4 and the thin plate portion 41 constitute the reaction well 2. A sample solution 46 and oil 47 are stored in the reaction well 2. When the reaction device 1 is disposed on the heater, heat is transmitted mainly from the lower surface side through the thin plate portion 41, and therefore, the thin plate portion 41 is preferably made of a material having a large heat transfer coefficient.

  As a reaction device used for nucleic acid analysis, a resin such as polypropylene is generally used. However, since the resin has a low heat transfer coefficient, it is desirable to use a metal having a higher heat transfer coefficient for the thin plate portion 41.

  Furthermore, among metals, it is desirable to use silver, copper, gold, aluminum, or the like having good thermal conductivity. In addition, heat transfer is conducted faster as the distance is shorter, and the loss is also smaller. Therefore, it is desirable that the thickness t1 of the thin plate portion 41 be as small as possible. When a metal is used for the thin plate portion 41, the sample may be contaminated. In that case, the adhesive layer 44 can also serve as a passivation layer. For example, an adhesive based on a silicone material can be used.

  Also in this case, it is desirable to make the thickness t4 of the adhesive layer 44 as small as possible in order to quickly transfer heat from the lower surface. Various materials can be used for the thick plate portion 42. For example, a resin such as polypropylene generally used for nucleic acid analysis can be used. Since heat transfer to the sample solution 46 and the oil 47 stored in the reaction well 2 is performed from the lower surface side, that is, the thin plate portion 41 and the adhesive layer 44 side, the heights t6 and t7 of the sample solution 46 and the oil 47 are It is desirable to make it as small as possible, and accordingly, it is desirable to make the thickness t2 of the thick plate portion 42 as small as possible. More specifically, the thickness t2 of the thick plate portion 42 is 1 mm or less, more preferably 0.5 mm or less.

  In addition, if the material of the thick plate portion is made of a material that does not easily transmit visible light, optical interference with adjacent wells can be blocked, and the S / N ratio in fluorescence intensity measurement can be improved. Is possible. For example, a resin such as polypropylene colored in black can be used. After the sample solution 46 and the oil 47 are placed in the reaction well 2, an evaporation prevention lid 43 is attached to the upper surface of the thick plate portion 42. Here, the oil 47 is used for preventing evaporation of the sample solution 46 until the lid 43 is attached. If the sample solution 46 can be stored in the reaction well 2 sufficiently quickly and the lid 43 can be attached, the oil 47 is not used. It doesn't matter.

  In the present invention, for the convenience of arranging a heater on the lower surface of the reaction device 1, the fluorescence intensity is measured through the upper surface side of the reaction device 1, that is, through the lid 43 and the adhesive layer 45. Therefore, the lid 43 and the adhesive layer 45 need to be a material that can transmit visible light. For these materials, for example, an adhesive polypropylene sheet having an adhesive layer in advance can be used. It is desirable to make the thicknesses t3 and t5 of the lid 43 and the adhesive layer 45 as small as possible so that the fluorescence is not attenuated.

  When the sample solution 46 becomes extremely small, it has a bottom structure for holding the sample solution 46 at a desired position in the reaction well 2. A recess 51 is formed in a partial region of the bottom surface 6 of the reaction well 2. The shape of the recess 51 as viewed from the top is circular in FIG. 10, but may be polygonal. Moreover, although the cross-sectional shape of the recessed part 51 is a semicircle in FIG. 10, it may be a square and other shapes. In FIG. 10, the recess 51 is arranged at the center of the bottom surface 6 of the reaction well 2, but the arrangement is not limited to the center as long as the irradiation detection of the optical system is possible. If the sample solution 46 is dropped into the recess 51, the sample solution 46 is held in the recess 51, so that the sample solution 46 can be disposed at a desired position.

  FIG. 11 is a top view showing the effect of the first means of the bottom structure for holding the sample solution 46. By forming the concave portion 51, the position of a small amount of the sample solution 46 held in all the reaction wells 2 can be disposed completely on the same circumference having the radius r1, and the optical system is securely fixed. Fluorescence measurement can be performed.

  FIG. 12 is a cross-sectional view showing the effect of the bottom surface structure means for holding the sample solution 46. Although the heat flow 61 from the thin plate portion 41 to the sample solution 46 is schematically shown, the contact area between the thin plate portion 41 and the sample solution 46 is increased by forming the concave portion 51 on the bottom surface 6 of the reaction well 2. Heat can be exchanged efficiently. Desirably, the volume formed by the bottom surface 6 of the reaction well 2 and the recess 51 is designed to be substantially the same as or slightly smaller than the volume of the sample solution 46. Furthermore, with this structure, it is possible to include all the sample solution 46 in the irradiation detection region 7 of the optical system. When the amount of sample solution becomes small, the amount of fluorescence decreases, but it is possible to improve the measurement sensitivity by capturing all the emitted fluorescence.

[Second Embodiment]
FIG. 13 (comparative example) is a cross-sectional view showing the behavior of the reaction device 100 shown in FIG. 7 when the temperature rises. The temperature of the reaction device 1 attached with the lid 430 at room temperature rises to about 95 ° C. during the PCR temperature cycle. In that case, the sample solution 460 is vaporized, and the internal pressure of the reaction well 200 increases. If the lid 430 does not have sufficient strength, the lid 430 is deformed by the increase in internal pressure, and in some cases, the adhesive layer 450 is peeled off from the thick plate portion 420, and a communication portion 480 is generated between the reaction wells. End up. When the communication part 480 is generated, contamination occurs between the communicating reaction wells, and accurate analysis cannot be performed. Therefore, in order to increase the strength, it is necessary to take measures such as making the lid 430 a highly rigid material or increasing the thickness t3 of the lid 430. In a reaction apparatus, in order to prevent contamination, a device once used for analysis is generally discarded. The above countermeasure is not a good measure because it leads to an increase in the manufacturing cost of the reaction device 1. Such a problem can also occur in FIG. 10 showing the above-described first embodiment of the present invention.

  FIG. 14 which shows this embodiment is a figure which shows the 2nd cross-sectional structure of the reaction device to which this invention is applied which solves this subject. In addition to the configuration shown in FIG. 10, a detachable weight 49 is added on the lid 43. Since the weight 49 is disposed on the upper surface of the lid 43, it does not directly contact the sample solution 46 and can be reused. In the present invention, for the convenience of arranging a heater on the lower surface of the reaction device 1, the fluorescence intensity is measured via the upper surface side of the reaction device 1, that is, through the weight 49, the lid 43 and the adhesive layer 45. Therefore, the weight 49 also needs to be a material that can transmit visible light. Quartz, glass, etc. can be used for this material. Quartz and glass generally have a high visible light transmittance, so that the thickness t8 of the weight 49 can be made relatively large although it is limited by the focal length of the optical system, and the inside of the reaction well 2 when the temperature rises. It will be able to withstand the increased pressure. Further, since the cross-sectional configuration of the present invention is realized by stacking different materials, there is a possibility that the reaction device may be distorted by the bimetal effect when rotating on a heater set at a plurality of temperatures. By adding the weight 49, the distortion of the reaction device is suppressed, and the adhesion between the reaction device and the heater is improved, so that heat transfer can be performed efficiently.

  Further, when a temperature cycle of about 95 ° C. to about 60 ° C. is applied to the reaction device of the present invention, the sample solution 46 vaporized at 95 ° C. may be condensed on the adhesive layer 45 when cooled to about 60 ° C. Condensed droplets cause light scattering when the fluorescence intensity is measured by the optical system 29, resulting in a decrease in signal intensity. As a method for solving this, condensation may be prevented by controlling the temperature of the weight 49 to about 95 to 100 ° C. As a method for controlling the temperature of the weight 49, heat may be applied by radiation from a remote heat source, or a surface 50 of the weight contacting the lid 43 may be made of a material that can transmit visible light such as ITO (Indium Tin Oxide). A heater may be formed.

  The cross-sectional configuration of the reaction device 1 shown in FIGS. 10 and 14 can be applied to any of the embodiments of the reaction device 1 shown in FIGS. 1, 4, 5, and 6. As shown, a plurality of reactors can be arranged in a reaction apparatus that performs PCR by contact rotation on a heater set at a plurality of temperatures.

  FIG. 15 is a perspective view showing a structure realized by a cross-sectional configuration of the reaction device of this example to which the present invention is applied. In FIG. 15, in order to display the structure in an easy-to-understand manner, the broken line portion is removed and shown. The reaction device 1 includes a thin plate portion 41, a thick plate portion 42, a lid 43, and a weight 49 from the lower surface side. Thin adhesive layers 44 and 45 are provided between the thin plate portion 41 and the thick plate portion 42 and between the thick plate portion 42 and the lid 43, respectively. The thick plate portion 42 is provided with a through hole, and constitutes the reaction well 2 having the thin plate portion 41 as a bottom surface. The reaction well 2 contains a sample solution 46 and oil 47 and is sealed with a lid 43. Further, the through hole 3 is provided in the lid 43, the thick plate portion 42, and the thin plate portion 41, and it is possible to fix the hole 3 by inserting a pin or the like when moving the reaction device. ing. In this embodiment, since the dimension in the longitudinal direction of the lid 43 is shorter than the dimension in the longitudinal direction of the thick plate portion 42, no hole 3 is provided in the lid 43, but the dimension in the longitudinal direction of the lid 43 is the thick plate portion 42. If the length is equal to the dimension in the longitudinal direction, the hole 3 may be provided in the lid 43 as well. All the reaction wells 2 are arranged on an arc having a radius r1, and a plurality of reaction devices 1 are provided in a reaction apparatus that performs PCR by contact rotation on a heater set at a plurality of temperatures as shown in FIG. When arranged, all the reaction wells are arranged on the circumference of the same radius r1.

[Third Embodiment]
FIG. 16 is a cross-sectional view showing the second means of the bottom structure for holding the sample solution 46 at a desired position in the reaction well 2 when the sample solution 46 becomes extremely small. A convex portion 52 is formed in a partial region of the bottom surface 6 of the reaction well 2. The shape of the projection 52 as viewed from the top is circular in FIG. 16, but may be polygonal. Moreover, although the cross-sectional shape of the convex part 52 is a triangle in FIG. 16, it may be a square or other shapes. In FIG. 16, the convex portion 52 is arranged at the center of the bottom surface 6 of the reaction well 2, but the arrangement is not limited to the center as long as the irradiation detection of the optical system is possible. If the sample solution 46 is dropped on the convex portion 52, the sample solution 46 is held in the concave portion 51, so that the sample solution 46 can be disposed at a desired position.

  FIG. 17 is a cross-sectional view showing the effect of the second means of the bottom structure for holding the sample solution 46. Although the heat flow 61 from the thin plate portion 41 to the sample solution 46 is schematically shown, the contact area between the thin plate portion 41 and the sample solution 46 is increased by forming the convex portion 52 on the bottom surface 6 of the reaction well 2. , Heat can be exchanged efficiently. Desirably, the shape of the convex part 52 is designed to be as close to a spherical surface as possible so that the surface area of the sample solution 46 and the contact part becomes as large as possible. Furthermore, with this structure, it is possible to include all the sample solution 46 in the irradiation detection region 7 of the optical system. When the amount of sample solution becomes small, the amount of fluorescence decreases, but it is possible to improve the measurement sensitivity by capturing all the emitted fluorescence.

  In any of the above embodiments, a hydrophobic treatment film may be formed in a region other than the concave portion or the convex portion that holds the sample. By forming such a film, the sample can be more reliably held at a predetermined position (concave or convex).

  As described in FIGS. 2 and 3, the reaction apparatus is equipped with the required number of reaction devices, and PCR is performed by contact rotation on a heater set at a plurality of temperatures, and the reaction is performed under the optical system 29. When the well 2 passes, the fluorescence intensity is measured. Even if there is a reaction device that has already started measurement, if there is a space on the heater, the reaction device can be inserted sequentially, and the reaction devices for which measurement has been completed can be sequentially discharged. Further, even if the sample solution 46 is in a very small amount, it is possible to reliably capture fluorescence by the optical system 29 by using the bottom structure described with reference to FIGS. 10, 14, and 16.

  As described above, by using the reaction device of the present invention, it is possible to measure the fluorescence intensity of all reaction wells with the optical system fixed, and realize a high-speed PCR apparatus that can continuously load / unload multiple reaction devices. It becomes possible to do.

DESCRIPTION OF SYMBOLS 1 Reaction device 2 Reaction well 3 Hole 4 Through-hole side wall 5 Reaction apparatus 6 Reaction well bottom face 7 Radiation detection area | regions 10-12 of an optical system, 14 Outer boundary 13 of reaction device The circular arc in which the outer boundary of reaction device and reaction well are formed Intersection 15 of the center 21-28 of the position where the reaction well is missing Heater 29 Optical system 41 Thin plate portion 42 Thick plate portion 43 Lid 44, 45 Adhesive layer 46 Sample solution 47 Oil 49 Weight 50 Surface of the weight contacting the lid 51 Reaction well Bottom concave portion 52 Reaction well bottom convex portion 61 Heat flow 100 Reaction device 200 Reaction well 400 Through-hole side wall 410 Thin plate portion 420 Thick plate portion 430 Lid 440,450 Adhesive layer 460 Sample solution 470 Oil 480 Interwell communication portion 600 Bottom surface of reaction well 700 Optical system irradiation detection area d1 Distance between adjacent reaction wells d2 Reaction weight The distance between the outer boundary and the reaction device outer boundary o The center r1 of the arc in which the reaction well is formed The radius r2-4 of the arc in which the reaction well is formed The radius t1 of the arc forming the outer boundary of the reaction device Thin plate thickness t2 Part thickness t3 Lid thickness t4, t5 Adhesive layer thickness t6 Sample solution height t7 Oil height t8 Weight thickness

Claims (19)

A reaction device used for temperature control of a solution containing a sample,
A solution storage section for storing the solution, and at least a thermally conductive first layer on the bottom side of the solution storage section;
A reaction device having a recess for holding a solution at the bottom of the solution storage unit. A reaction device used for temperature control of a solution containing a sample,
A solution storage section for storing the solution, and at least a thermally conductive first layer on the bottom side of the solution storage section;
A reaction device having a convex portion for holding a solution at a bottom portion of the solution storage portion. The reaction device according to claim 1 or 2,
A reaction device comprising a second layer that constitutes a side wall of the solution storage section above the first layer. The reaction device according to claim 3,
A plurality of solution storage units are provided,
The reaction device according to claim 2, wherein the second layer prevents optical interference between adjacent solution storage units. The reaction device according to claim 3,
A reaction device comprising a third layer on an upper portion of the second layer for preventing evaporation of the solution stored in the solution storage section. The reaction device according to claim 5, wherein
A reaction device comprising a fourth layer as a weight for preventing the third layer from being bent on an upper portion of the third layer. The reaction device according to claim 3,
A reaction device comprising an adhesive layer between the first layer and the second layer. The reaction device according to claim 1,
A reaction device, characterized in that a hydrophobic film is provided in a portion other than the concave portion on the bottom surface of the solution storage portion. The reaction device according to claim 2,
A reaction device, characterized in that a hydrophobic film is provided in a portion other than the convex portion on the bottom surface of the solution storage portion. The reaction device according to claim 5, wherein
A reaction device characterized in that the third layer is made of a material capable of transmitting visible light. The reaction device according to claim 6,
The reaction device according to claim 4, wherein the fourth layer is made of a material capable of transmitting visible light. A reaction device including a solution storage unit that stores a solution containing a sample, and a thermally conductive first layer at least on the bottom side of the solution storage unit;
A temperature adjustment unit for adjusting the temperature of the solution in the solution storage unit,
It is equipped with multiple reaction devices and is detachable.
A reactor having a recess for holding a solution at the bottom of the solution storage unit. A reaction device including a solution storage unit that stores a solution containing a sample, and a thermally conductive first layer at least on the bottom side of the solution storage unit;
A temperature adjustment unit for adjusting the temperature of the solution in the solution storage unit,
It is equipped with multiple reaction devices and is detachable.
A reaction apparatus having a convex part for holding a solution at a bottom part of the solution storage part. A reactor according to claim 12 or 13,
The temperature adjusting unit comprises a plurality of temperature control units that can be individually temperature-adjusted and are adjacent to each other. 15. A reactor according to claim 14, wherein
A reaction apparatus comprising a drive unit for moving a reaction device, wherein the reaction device is moved between temperature control units by the drive unit. A reactor according to claim 12 or 13,
A reaction device characterized in that the reaction device is movable on the temperature adjustment unit. A reactor according to claim 12 or 13,
The reaction device is provided with a plurality of solution storage portions, and each solution storage portion is arranged in an arc shape. A reactor according to claim 17,
The reaction apparatus characterized in that the planar shape of the solution storage part is an oval shape curved along the arc shape. A reactor according to claim 12 or 13,
A reaction apparatus comprising an optical unit for irradiating excitation light and observing fluorescence.