WO2006132324A1 - Reaction container and reaction apparatus employing the same - Google Patents

Abstract

A reaction container (110) comprising a DNA microarray (111), a upper cylinder (112), a lower cylinder (113) and an elastic member (114). The DNA microarray (111) is formed by immobilizing two or more nucleic acid probes on a porous substrate, through which a sample solution containing a biological substance to be tested can permeate. The upper cylinder (112) and the lower cylinder (113) are connected to each other in such a manner that the DNA microarray (111) is sandwiched between them. The DNA microarray (111) is retained in such a manner that it is placed across the interior space of a cylindrical body formed with the upper cylinder (112) and the lower cylinder (113). The elastic member (114) has a dome-like shape and serves to seal the open end of the lower cylinder (113).

Description

 Specification

 Reaction vessel and reaction apparatus using the same

 Technical field

 TECHNICAL FIELD [0001] The present invention relates to a reaction container used for inspection of a biological substance.

 Background art

 Recently, a novel genetic testing method using a test substrate V in which single-stranded DNA is regularly arranged on a substrate such as a semiconductor or glass, a so-called DNA chip or a DNA microarray has been developed. This test method has the IJ point that multiple genes can be tested simultaneously.

 [0003] Regarding such a gene testing method, Japanese Patent Publication No. 10-505410 discloses a technique related to a reaction chamber that accommodates a DNA microarray. In the embodiment disclosed in Japanese Patent Publication No. 10-5054 10, it is attached to a main body having a cavity using a substrate force having a probe array, for example, an adhesive. In addition, the body has an inlet that allows fluid to flow into (and through) the cavity to detect the gene by a hybridization reaction between the sample and the probe. A medium is formed.

 [0004] Patent No. 3488465 uses a fine glass tube integrated substrate or a porous silicon substrate as the substrate on which the probe array is formed, thereby reducing the time required for the hybridization reaction and detecting it. A technique for improving sensitivity is disclosed. Furthermore, a reaction chamber using a Delrin O-ring is disclosed as an embodiment of a reaction apparatus for carrying out a hybridization reaction using a DNA microarray using this porous substrate.

 [0005] These reaction containers and reaction chambers have a structure that can promote a hybridization reaction by flowing a sample solution to a DNA microarray.

 Disclosure of the invention

[0006] The reaction chamber disclosed in Japanese Patent Publication No. 10-505410 has a structure in which a pipe for flowing the sample solution is directly connected. There is concern about contamination of the device itself. For this reason, when carrying out repeated inspections, it is necessary to clean the inside of the reaction apparatus, and the apparatus becomes heavy.

 [0007] On the other hand, in the technique disclosed in Japanese Patent No. 3488465, since the reaction chamber 1 is fixedly connected to the pressure source, it is difficult to automatically process a large number of DNA microarrays.

 [0008] The present invention has been made in consideration of such a situation, and an object of the present invention is to provide a reaction vessel that does not require cleaning of the reaction apparatus.

 [0009] The reaction container of the present invention seals a reaction substrate having a probe that reacts with a biological substance, a channel in which the reaction substrate is exposed, and one end of the channel. A sample solution in which a biological substance is dissolved from another end of the flow path that is sealed, and is present in the flow path. The sample solution to flow flows with respect to the reaction substrate according to the elastic deformation of the elastic member.

 Brief Description of Drawings

 [0010] FIG. 1 shows a reaction vessel in a first embodiment of the present invention.

 FIG. 2 shows a reaction apparatus according to the first embodiment of the present invention.

 FIG. 3 shows a reaction vessel in the second embodiment of the present invention.

 FIG. 4 shows a reaction apparatus in a second embodiment of the present invention.

 FIG. 5 shows a genetic test apparatus according to the third embodiment of the present invention.

 FIG. 6 shows a modification of the genetic testing device in the third embodiment of the present invention.

 FIG. 7 shows a reaction vessel in the fourth embodiment of the present invention.

 FIG. 8 shows a reaction apparatus in a fourth embodiment of the present invention.

 FIG. 9 shows a reaction vessel in a fifth embodiment of the present invention.

 FIG. 10 shows a cross section of the reaction vessel along the line XX in FIG.

 [FIG. 11] FIG. 11 shows a reaction apparatus according to a fifth embodiment of the present invention! / Speak.

 FIG. 12 shows a modification of the reaction vessel in the second embodiment of the present invention.

FIG. 13 shows a modification of the reaction apparatus in the second embodiment of the present invention. BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, embodiments of the present invention in the case of detecting a nucleic acid as an example of a biological substance will be described with reference to the drawings. Here, a nucleic acid is taken as an example, but the present invention is not limited to this, and it may be a protein or other biological substance.

 <First Embodiment>

 [Constitution]

 FIG. 1 shows a reaction vessel in the first embodiment of the present invention. The reaction container 110 includes a DNA microarray 111, an upper cylinder 112, a lower cylinder 113, and an elastic member 114. The DNA microarray 111 is a reaction substrate having a probe that reacts with a biological substance, and is prepared by immobilizing a plurality of nucleic acid probes in a plurality of regions 1 15 on a substrate that transmits a solution such as a porous substrate. It can permeate the sample solution containing the biological material to be examined. The upper cylinder 112 has a flange portion 112a at the upper end. The upper cylinder 112 and the lower cylinder 113 are joined to each other with the DNA microarray 111 interposed therebetween, and the inner space forms a single cylinder that defines a flow path through which a gas or liquid passes. The DNA microarray 111 is exposed in the inner space of the cylinder composed of the upper cylinder 112 and the lower cylinder 113, is held so as to cross the inner space, and extends substantially perpendicular to the axis of the cylinder. As will be described later, the sample solution is reciprocated substantially parallel to the axis of the cylinder. That is, the DNA microarray 111 extends substantially perpendicular to the flow direction of the sample solution. The elastic member 114 has a dome shape and seals the open end of the lower cylinder 113.

 FIG. 2 shows a reaction apparatus in the first embodiment of the present invention. The reaction apparatus 120 includes a reaction container housing part 130 that houses the reaction container 110 and a drive part 140 that deforms the elastic member 114 of the reaction container 110.

 [0014] The reaction vessel storage unit 130 includes a storage unit body 131 for receiving the reaction vessel 110, a Peltier element 132 for adjusting the temperature of the reaction vessel 110, and a temperature sensor 133 for measuring the temperature of the reaction vessel 110, A lid 134 for fixing the reaction vessel 110, and a temperature control device 135 for controlling the Peltier element 132 based on information from the temperature sensor 133.

The storage unit main body 131 has a space in which the reaction vessel 110 is inserted and a step portion that receives the flange portion 112a of the reaction vessel 110. The reaction vessel 110 inserted into the container body 131 is The flange portion 112 a is supported by the housing portion main body 131 by being received by the step portion of the housing portion main body 131. The lid 134 can be opened and closed with respect to the housing part body 131. The reaction vessel 110 accommodated in the accommodating portion main body 131 is fixed by closing the lid 134. The lid 134 has a through hole 134a, and the sample solution S can be dispensed into the reaction vessel 110 accommodated in the accommodating portion main body 131 through the through hole 134a.

 The drive unit 140 includes a piston 141 for pressing the elastic member 114, a crankshaft 143 rotated by a motor, and a connecting rod 142 connecting the piston 141 and the crankshaft 143. Thus, the rotational motion of the crankshaft 143 is converted into the reciprocating linear motion of the piston 141.

 [0017] [Action]

 Hereinafter, the operation of the reaction vessel 110 and the reaction device 120 of the present embodiment will be described with respect to the reaction device 12.

The operation will be described according to the operation procedure 0.

(Procedure 1) The reaction vessel 110 is set in the reaction vessel housing part 130.

(Procedure 2) The lid 134 is closed and the reaction vessel 110 is fixed to the reaction vessel housing part 130.

[0020] (Procedure 3) The crankshaft 143 is rotated to move the piston 141 upward as shown on the left side of FIG. As a result, the elastic member 114 is crushed and the volume of the space below the DNA microarray 111 is reduced.

[0021] (Procedure 4) Using a pipette, the sample solution S containing the nucleic acid sample to be measured, which is preliminarily labeled with a fluorescent substance by a known technique, is separated on the DNA microarray 111 from the through-hole 134a of the lid 134. Note.

(Procedure 5) Based on the signal from the temperature sensor 133, the temperature control device 135 controls the Bellech element 132 to adjust the reaction vessel 110 to a temperature suitable for the hybridization reaction.

 [Step 6] Rotate the crankshaft 143 to move the piston 141 downward as shown on the right side of FIG. As a result, the elastic member 114 returns to its original shape, and the volume of the space below the DNA microarray 111 increases. As the volume increases, the sample solution S passes through the DNA microarray 111 and moves to the lower side of the DNA microarray 111.

(Procedure 7) Rotate the crankshaft 143 and lift the piston 141 as shown on the left side of FIG. Move towards. As a result, the elastic member 114 is crushed and the volume of the space below the DNA microarray 111 is reduced. As the volume decreases, the sample solution S passes through the DNA microarray 111 and moves to the upper side of the DNA microarray 111.

 [Step 8] Repeat Step 6 and Step 7. As a result, the sample solution S repeatedly permeates through the DN A microarray 111 and flows back and forth. That is, the sample solution S is fluidized by pressing and releasing by the piston 141, that is, by mechanical means. This facilitates the post-hybridization reaction.

 (Procedure 9) After the predetermined reaction is completed, the lid 134 is opened, and the reaction vessel 110 is taken out from the reaction vessel housing part 130.

 [0027] (Procedure 10) Do not show the extracted DNA microarray 111 image! Fluorescence microscope and CC

D or acquired by a fluorescent image acquisition device such as a laser scanning microscope.

(Procedure 11) The acquired fluorescence image is analyzed to obtain genetic information of the sample.

[0029] [Effect]

 In the present embodiment, the sample solution S in the reaction vessel 110 is fluidized by deformation of the elastic member 114 provided in the reaction vessel 110. For this reason, the sample solution S is reciprocated while the solution storage space in the reaction vessel 110 is isolated from the drive unit 140. Therefore, since the reactor 120 is not contaminated by the sample solution in the reaction vessel 110, there is no need for cleaning.

[0030] [Modification]

 The reaction vessel 110 shown in FIGS. 1 and 2 has a dome-shaped elastic member 114, but the shape of the elastic member is not limited to this, and the reaction vessel has, for example, a dropper-type elastic member. It may be. In the reaction container 110 shown in FIGS. 1 and 2 in order to flow the sample solution S, the force by which the elastic member 114 is pressed in parallel with the attaching / detaching direction of the reaction container 110 to the reaction container housing part 130. In the container, a dropper-type elastic member may be pressed in a direction perpendicular to the attaching / detaching direction of the reaction container. As a result, the force required for positioning the reaction vessel is small, and the requirement for the reaction vessel fixing mechanism in the reaction apparatus is reduced.

[0031] <Second embodiment>

[Constitution] FIG. 3 shows a reaction vessel in the second embodiment of the present invention. In FIG. 3, members indicated by the same reference numerals as those shown in FIG. 1 are the same members, and detailed description thereof is omitted. The reaction vessel 210 includes a bellows-like elastic member 214 and a magnetic metal plate 215 provided on the elastic member 214 instead of the elastic member 114 of the reaction vessel 110 shown in FIG. That is, the reaction vessel 210 has a DNA microarray 111, an upper cylinder 112, a lower cylinder 113, an elastic member 214, and a magnetic metal plate 215. The elastic member 214 seals the opening end of the lower cylinder 113, and the magnetic metal plate 215 is an iron plate, for example, and is fixed to the bottom surface of the elastic member 214.

 FIG. 4 shows a reaction apparatus in the second embodiment of the present invention. In FIG. 4, members indicated by the same reference numerals as those shown in FIG. 2 are similar members, and detailed description thereof is omitted. The reaction device 220 includes a drive unit 240 that deforms the elastic member 114 of the reaction vessel 110 in place of the drive unit 140 of the reaction device 120 shown in FIG. In addition to the piston 141, the crankshaft 143, and the connecting rod 142, the drive unit 240 includes an electromagnet 244 provided on the piston 141. The electromagnet 244 is located at the upper end of the piston 141, and adsorbs the magnetic metal plate 215 as necessary.

 [0033] [Action]

 The operation procedure of the reaction apparatus 220 in this embodiment is the same as the procedure in the first embodiment. However, in this embodiment, the following steps are added to the procedure of the first embodiment.

 (Procedure 2-1) Following the procedure 2, the electromagnet 244 is energized to connect the bellows-like elastic member 214 of the reaction vessel 210 and the piston 141.

 (Procedure 8—1) Following step 8, the electromagnet 244 is de-energized and the connection between the bellows-like elastic member 214 of the reaction vessel and the piston 141 is disconnected.

 [0036] [Effect]

Also in the present embodiment, the sample solution S is reciprocated while the solution storage space in the reaction vessel 210 is isolated from the drive unit 240. Therefore, since the reactor 220 is not contaminated by the sample solution in the reaction vessel 110, there is no need for cleaning. Furthermore, since the elastic member 214 is a bellows, the volume of the space below the DNA microarray 111 can be changed greatly. [0037] [First Modification]

 The reaction vessel 210 shown in FIG. 3 has the magnetic metal plate 215 below the elastic member 214, but may have a magnet instead. That is, the reaction vessel 210 may have a magnet provided on the elastic member 214 instead of the magnetic metal plate 215.

[0038] [Second modification]

 Further, in the reactor 220 shown in FIG. 4, the force connecting the elastic member 214 and the piston 141 by utilizing the adsorption of the electromagnet 244 and the magnetic metal plate 215, as shown in FIGS. The elastic member 214 and the piston 141 may be connected using the adsorption force. That is, the reaction vessel 210 may have a suction cup 216 provided on the elastic member 214 instead of the magnetic metal plate 215. In this case, in order to disconnect the elastic member 214 and the piston 141, the piston 141 is provided with an electromagnetic valve 246 that opens to atmospheric pressure.

[0039] <Third embodiment>

 [Constitution]

 FIG. 5 shows a genetic test apparatus according to the third embodiment of the present invention. The genetic test apparatus of the present embodiment uses the reaction vessel 110 of the first embodiment. In FIG. 5, members indicated by the same reference numerals as those shown in FIGS. 1 and 2 are the same members, and detailed description thereof will be omitted.

[0040] The genetic testing apparatus 300 stocks a turntable 301 for transporting the reaction vessel 110, a plurality of drive units 140 for flowing the sample solution in the reaction vessel 110, and a plurality of reaction vessels 110. A reaction container storage section 302, a reaction container operation arm 303 for operating the reaction container 110, a sample rack 304 for storing a container in which a sample solution containing a nucleic acid sample is dispensed, and a sample stored in the sample rack 304 A sample dispensing mechanism 305 for dispensing the solution into the reaction vessel 110, a measurement unit 306 for taking a fluorescent image of the DNA microarray 111 in the reaction vessel 110, and a control device 307 for controlling the entire device. Have it!

[0041] The turntable 301 has a plurality of reaction container storage portions 130 for storing the reaction container 110, and the reaction container 110 is placed in a plurality of stop positions (reaction container mounting position 309, sample solution dispensing position 310, a plurality of Reaction execution position 311, image acquisition position 312, reaction container disposal position 313) Transport. The reaction container housing part 130 is substantially the same as that described in the first embodiment. The reaction vessel operation arm 303 takes out the reaction vessel 110 from the reaction vessel storage unit 302 and supplies it to the reaction vessel storage unit 130, and takes out the reaction vessel 110 from the reaction vessel storage unit 302 at the reaction vessel disposal position 313. Dispose of in the reaction vessel disposal port 308. The plurality of drive units 140 are respectively disposed at the plurality of stop positions described above. Each drive unit 140 is the same as that described in the first embodiment. The sample dispensing mechanism 305 dispenses the sample solution into the reaction container 110 through the through-hole 1 34a of the lid 134 of the reaction container housing part 130, and the measurement unit 306 acquires an image of the DNA microarray 111 in the reaction container 110. .

[0042] [Action]

 Hereinafter, the operation of the genetic test apparatus 300 of the present embodiment will be described according to the operation procedure and process. Here, the procedure means work performed by the user, and the process means work performed by the apparatus.

(Procedure 1) A plurality of reaction vessels 110 are set in the reaction vessel storage section 302.

(Procedure 2) In the sample rack 304, a sample solution containing a nucleic acid sample to be measured and labeled with a fluorescent substance in advance by a known technique is set in a state of being dispensed into a container.

(Procedure 3) Instruct the genetic testing apparatus 300 to start testing.

 (Step 1) The reaction vessel operating arm 303 moves the reaction vessel 11 from the reaction vessel storage section 302.

0 is taken out and set in the reaction container housing part 130 at the reaction container mounting position 309.

(Step 2) The lid 134 of the reaction container housing part 130 at the reaction container mounting position 309 is closed.

(Step 3) The turntable 301 is rotated to transport the reaction vessel 110 to the sample solution dispensing position 310.

 (Step 4) The piston 141 of the drive unit 140 at the sample solution dispensing position 310 is moved upward to crush the elastic member 114 of the reaction vessel 110.

(Step 5) The sample solution stored in the sample rack 304 by the sample dispensing mechanism 305 is dropped into the reaction vessel 110.

(Step 6) The turntable 301 is rotated to transport the reaction vessel 110 to the reaction execution position 311.

[0052] (Step 7) The temperature of the reaction container housing part 130 at the reaction execution position 311 is hybridized. The temperature is controlled to be suitable for the Chillon reaction.

 (Step 8) The sample solution is moved to the lower side of the DNA microarray 111 by moving the piston 141 of the drive unit 140 at the reaction execution position 311 downward to return the elastic member 114 to its original shape. Furthermore, the sample solution is moved to the upper side of the DNA microarray 111 by moving the piston 141 of the drive unit 140 upward to crush the elastic member 114. By repeating this operation, the sample solution is reciprocated to repeatedly pass through the DNA microarray 111.

 (Step 9) While the turntable 301 is rotated to transport the reaction vessel 110, step 8 is repeatedly executed at each reaction execution position 311.

 (Step 10) After a predetermined reaction time has elapsed, the reaction vessel 110 is placed at the image acquisition position 312 below the measurement unit 306.

 (Step 11) The piston 141 of the drive unit 140 at the image acquisition position 312 is moved downward to move the sample solution to the lower side of the DNA microarray 111.

 (Step 12) An image of the DNA microarray 111 in the reaction vessel 110 is acquired by the measurement unit 306.

 (Step 13) Rotate the turntable 301 to place the reaction vessel 110 that has been photographed at the reaction vessel disposal position 313.

(Step 14) Open the lid 134 of the reaction container housing part 130 at the reaction container disposal position 313.

(Step 15) The reaction vessel 11 is moved from the reaction vessel housing section 130 by the reaction vessel operation arm 303.

Take out 0 and discard it in the reaction container waste outlet 308.

(Step 16) The fluorescence image acquired by the measurement unit 306 is analyzed to obtain the gene information of the sample.

 [0062] In the description of Step 1 to Step 16 described above, attention is given to one reaction vessel 110 in order to explain the processing flow of the reaction vessel 110, but actually, a plurality of reaction vessels 110 are included. One by one is continuously supplied to the turntable 301, and Step 1 to Step 16 are sequentially and sequentially repeated for the reaction vessel 110 supplied to the turntable 301. As a result, a large number of reaction vessels 110 are automatically processed within a short time.

[0063] [Effect] According to the genetic test apparatus 300 of the present embodiment, from the gene detection reaction to the acquisition of image data is automatically performed simply by the user setting the reaction container and the sample solution. Similarly to the first embodiment, the sample solution is reciprocated while the solution storage space in the reaction vessel 110 is isolated from the drive unit 140. Therefore, since the genetic test apparatus 300 is not contaminated by the sample solution in the reaction vessel 110, there is no need for cleaning.

 [0064] [First Modification]

 The genetic test apparatus 300 shown in FIG. 5 is configured to use the reaction container 110 and the reaction apparatus 120 of the first embodiment, but may be configured to use the reaction container 210 and the reaction apparatus 220 of the second embodiment. Good. That is, the genetic test apparatus 300 may include the drive unit 240 of the second embodiment instead of the drive unit 140, and the reaction vessel 210 of the second embodiment may be used instead of the reaction vessel 110.

 [0065] [Second modification]

 5 has a plurality of independent drive units 140 at the stop position of the turntable 301 in order to press the elastic member 114 of the reaction vessel 110. As shown in FIG. 6, instead of the plurality of drive units 140, a piston 141 provided for each stop position of the turntable 301, a cam follower 314 connected to the piston 141, and the turntable 301 are coaxial. And a cam 315 that rotates. The cam 315 has a cam surface whose height changes periodically, and the cam follower 314 moves the piston 141 up and down according to the rotation of the cam 315. In this modification, the plurality of pistons 141 are moved up and down by a single rotational power source such as a motor, so that the device configuration is simplified.

 [0066] <Fourth embodiment>

 [Constitution]

FIG. 7 shows a reaction vessel in the fourth embodiment of the present invention. In FIG. 7, the members indicated by the same reference numerals as those shown in FIG. 3 are the same members, and the details LV and description thereof are omitted. The reaction vessel 410 has a DNA microarray 111, an upper cylinder 112, a lower cylinder 113, and an elastic member 214. The elastic member 214 has a bellows shape and seals the open end of the lower cylinder 113. ing. That is, the reaction vessel 410 has a configuration in which the magnetic metal plate 215 is omitted from the reaction vessel 210 shown in FIG.

 FIG. 8 shows a reaction apparatus according to the fourth embodiment of the present invention. The reaction apparatus 420 includes a reaction container housing part 430 for housing the reaction container 410 and a drive part 440 for deforming the elastic member 214 of the reaction container 410.

 [0068] The reaction vessel storage unit 430 includes a storage unit body 431 that receives the reaction vessel 410, a Peltier element 432 that adjusts the temperature of the reaction vessel 410, and a temperature sensor 433 that measures the temperature of the reaction vessel 410. And a lid 434 for fixing the reaction vessel 410 and a temperature control device 435 for controlling the Peltier element 432 based on the information of the temperature sensor 433.

 [0069] The container main body 431 includes a space into which the reaction vessel 410 is inserted and a step portion that receives the flange portion 112a of the reaction vessel 410 via the O-ring 436. The reaction vessel 410 inserted into the container main body 431 is supported by the container main body 431 when the flange portion 112a is received by the step portion of the container main body 431 via the O-ring 436. The lid 434 can be opened and closed with respect to the housing main body 431. The reaction container 410 accommodated in the accommodating body 431 is fixed by closing the lid 434. The space between the reaction vessel 410 and the container main body 431 is kept airtight by an O-ring 436. The lid 434 has a through hole 434a, and the sample solution S can be dispensed into the reaction container 410 accommodated in the accommodating portion main body 431 through the through hole 434a. The accommodating portion main body 431 has a cylindrical projecting portion 431a at the bottom, and the inner space of the accommodating portion main body 431 communicates with the outside via the cylindrical projecting portion 431a.

 [0070] The drive unit 440 includes a syringe pump 441 and a tube 446 that connects the distal end portion of the syringe pump 441 and the cylindrical projecting portion 43la of the housing main body 431.

 [0071] [Action]

 (Procedure 1) Set the reaction vessel 410 in the reaction vessel storage section 430.

 (Procedure 2) The lid 434 is closed and the reaction vessel 410 is fixed to the reaction vessel housing part 430.

(Procedure 3) The piston 441a is driven upward as shown on the left side of FIG. As a result, the inner space of the container main body 431 is pressurized, the elastic member 214 is crushed, and the volume of the space below the DNA microarray 111 is reduced. [0074] (Procedure 4) Using a pipette, the sample solution is dispensed onto the DNA microarray 111 from the through hole 434a of the lid 434.

(Procedure 5) Based on the signal from the temperature sensor 133, the temperature control device 435 controls the Bellech element 132 to adjust the reaction vessel 410 to a temperature suitable for the hybridization reaction.

 (Procedure 6) The piston 441a is driven downward as shown on the right side of FIG. As a result, the inner space of the container main body 431 is decompressed, the elastic member 214 returns to its original shape, and the volume of the space below the DNA microarray 111 increases. As the volume increases, the sample solution S passes through the DNA microarray 111 and moves to the lower side of the DNA microarray 111.

 (Procedure 7) The piston 441a is driven upward as shown on the left side of FIG. As a result, the inner space of the container main body 431 is pressurized, the elastic member 214 is crushed, and the volume of the space below the DNA microarray 111 is reduced. As the volume decreases, the sample solution S passes through the DNA microarray 111 and moves to the upper side of the DNA microarray 111.

 [0078] (Step 8) Repeat Step 6 and Step 7. As a result, the sample solution S repeatedly permeates through the DN A microarray 111 and flows back and forth. That is, the sample solution S is flowed by pressurization and depressurization via air, that is, by fluid means. As a result, no reaction after the hybridization reaction is promoted.

 (Procedure 9) After the predetermined reaction is completed, the lid 434 is opened, and the reaction vessel 410 is taken out from the reaction vessel storage unit 430.

 [0080] (Procedure 10) Do not show the image of the extracted DNA microarray 111 !, fluorescence microscope and CC

D or acquired by a fluorescent image acquisition device such as a laser scanning microscope.

(Procedure 11) The acquired fluorescence image is analyzed to obtain genetic information of the sample.

[0082] [Effect]

In the present embodiment, the sample solution S in the reaction vessel 410 flows due to deformation of the elastic member 214 provided in the reaction vessel 410. For this reason, the sample solution S is reciprocated while the solution storage space in the reaction vessel 410 is isolated from the drive unit 440. Therefore, the reactor 420 is not contaminated by the sample solution in the reaction vessel 110. No need for cleaning. Further, since the elastic member 214 is deformed by fluid means, the number of parts of the reaction vessel 410 can be reduced, and the reaction vessel 410 can be easily manufactured.

 [0083] <Fifth embodiment>

 Here, an embodiment in the case of a DNA microarray using a substrate, in which the sample solution does not penetrate and circulate, will be described with reference to FIGS. 9, 10, and 11. FIG.

 [0084] [Configuration]

 FIG. 9 shows a reaction vessel in the fifth embodiment of the present invention, and FIG. 10 shows a cross section of the reaction vessel along the line XX in FIG. The reaction vessel 510 has a DNA microarray 511, a flow path member 512, an elastic member 514, a lower housing 515, and an upper housing 516. The DNA microarray 511 is a reaction substrate having a probe that reacts with a biological substance, does not permeate a solution such as a glass substrate, and is produced by immobilizing a plurality of nucleic acid probes on a substrate in a plurality of regions 517. And cannot pass through the sample solution. The flow path member 512 has two through holes 512a and 512b and a groove 512c extending between the two through holes 512a and 512b. The flow channel member 512 cooperates with the DNA microarray 511 to allow a gas or liquid to flow therethrough. Define the road. That is, the nucleic acid probe immobilized on the DNA microarray 511 is exposed in the channel and extends along the groove 512c. As will be described later, the sample solution is reciprocated in the groove 512c. That is, the nucleic acid probe immobilized on the DNA microarray 511 extends almost in parallel with the flow direction of the sample solution. The elastic member 514 has a dome shape, and seals the through hole 512b of the flow path member 512 that hits one end of the flow path. The lower casing 515 and the upper casing 516 are joined to each other, and are held by sandwiching the DNA microarray 511 and the flow path member 512. The flow path member 512 and the elastic member 514 may be integrally formed of an elastic material.

 FIG. 11 shows a reaction apparatus according to the fifth embodiment of the present invention. The reaction apparatus 520 includes a reaction container storage unit 530 that stores the reaction container 510 and a drive unit 540 that deforms the elastic member 514 of the reaction container 510.

[0086] The reaction container housing unit 530 includes a housing body 531 that receives the reaction container 510, a Peltier element 532 for adjusting the temperature of the reaction container 510, and a temperature sensor 533 for measuring the temperature of the reaction container 510. A lid 534 for fixing the reaction vessel 510 and a temperature sensor And a temperature control device 535 for controlling the Peltier element 532 based on 533 information.

 The accommodating portion main body 531 has a recess into which the reaction vessel 510 is inserted. In the reaction vessel 510 inserted into the housing main body 531, the DNA microarray 511 is supported in contact with the bottom of the recess. The lid 534 can be opened and closed with respect to the housing main body 531. The reaction vessel 510 accommodated in the accommodating body 531 is fixed by closing the lid 534. The lid 534 has two through holes 534a and 534b. The through hole 534a is located above the through hole 512a of the reaction vessel 510, and the sample solution S is divided into the through hole 512a of the reaction vessel 510 through the through hole 534a. The through hole 534b is located above the elastic member 514 of the reaction vessel 510.

 [0088] The drive unit 540 has a piston 541 for pressing the elastic member 514 and an actuator 542 for moving the piston 541 up and down!

[0089] [Action]

 The operation of the reaction vessel 510 and the reaction apparatus 520 of the present embodiment will be described according to the operation procedure of the reaction apparatus 520.

(Procedure 1) The reaction vessel 510 is set in the reaction vessel storage unit 530.

(Procedure 2) The lid 534 is closed and the reaction vessel 510 is fixed to the reaction vessel housing part 530.

(Procedure 3) The actuator 542 is driven to move the piston 541 downward as shown in the lower side of FIG. As a result, the elastic member 514 is crushed and the volume of the space inside the elastic member 514 is reduced.

[0093] (Procedure 4) Dispense the sample solution into the through-hole 512a of the reaction vessel 510 using a pipette.

(Procedure 5) Based on the signal from the temperature sensor 533, the temperature control device 535 controls the Verge element 532 to adjust the reaction vessel 510 to a temperature suitable for the hybridization reaction.

(Procedure 6) The actuator 542 is driven to move the piston 541 upward as shown in the upper side of FIG. As a result, the elastic member 514 returns to its original shape, and the volume of the space inside the elastic member 514 increases. As the volume increases, the sample solution moves to the right in groove 512c. And pass through the probe region of the DNA microarray 511.

(Procedure 7) The actuator 542 is driven to move the piston 541 downward as shown in the lower side of FIG. As a result, the elastic member 514 is crushed, and the volume of the space inside the elastic member 514 is reduced. As the volume decreases, the sample solution moves to the left in the groove 512c and passes through the probe region of the DNA microarray 511.

[0097] (Step 8) Repeat Step 6 and Step 7. As a result, the sample solution reciprocates in the groove 512c and repeatedly passes through the probe region of the DNA microarray 511. As a result, the post-nozzle reaction is promoted.

(Procedure 9) After the predetermined reaction is completed, the lid 534 is opened, and the reaction vessel 510 is taken out from the reaction vessel storage unit 530.

 [0099] (Procedure 10) Fluorescent microscope and CC (not shown) showing the extracted DNA microarray 511 image

D or acquired by a fluorescent image acquisition device such as a laser scanning microscope.

[0100] (Procedure 11) Obtain the genetic information of the sample by analyzing the acquired fluorescence image.

[0101] [Effect]

 In the present embodiment, the sample solution in the reaction vessel 510 is fluidized by deformation of the elastic member 514 provided in the reaction vessel 510. Therefore, the sample solution is reciprocated while the solution storage space in the reaction vessel 510 is isolated from the drive unit 540. Therefore, the reaction device 520 is not contaminated by the sample solution in the reaction vessel 510, and thus does not need to be cleaned. In addition, since the reaction vessel housing portion 530 is on the extension line in the pressing direction by the piston 541, the reaction vessel 510 can be easily held.

[0102] So far, the embodiments of the present invention have been described with reference to the drawings. The present invention is not limited to these embodiments, and various modifications and changes can be made without departing from the scope of the present invention. May be.

 Industrial applicability

[0103] According to the present invention, there is provided a reaction vessel that does not require cleaning of the reaction apparatus!

Claims

The scope of the claims  [1] a reaction substrate (111; 511) having a probe that reacts with a biological substance;  A flow path in which the reaction substrate is exposed;  An elastic member (114; 214; 514) that seals one end of the flow path, and is sealed with the flow path. The liquid is supplied into the flow path, and the sample solution existing in the flow path is the elastic member. (114; 214; 514) A reaction vessel (110; 210; 410; 510), which flows relative to the reaction substrate (111; 511) in accordance with elastic deformation. [2] The reaction vessel (110; 210; 410) according to claim 1, wherein the reaction substrate (111) can permeate the sample solution. [3] The reaction vessel (110; 210; 410) according to claim 2, wherein the reaction substrate (111) includes a porous substrate. [4] The reaction vessel (110; 210; 410) according to claim 2, wherein the reaction substrate (111) extends substantially perpendicular to the flow direction of the sample solution. [5] It has a cylinder (112, 113) that defines the flow path, and the reaction substrate (111) is held by the cylinder (112, 113) so as to cross the flow path, and the elastic member ( 114; 214) The reaction container (110; 210; 410) according to claim 4, wherein one end of the cylindrical body (112, 113) is sealed. 6. The reaction vessel according to claim 5, wherein the elastic member (214) has a bellows shape. 210; 410).  [7] The reaction vessel (210; 410) according to claim 5, further comprising a suction cup (216) provided on the elastic member (214). [8] The reaction vessel (210; 410) according to claim 5, further comprising a magnetic metal plate (215) provided on the elastic member (214). [9] The method further comprises a magnet provided on the elastic member (214). 5. The reaction vessel (210; 410) according to 5. [10] The reaction vessel (510) of claim 1, wherein the reaction substrate (511) cannot pass through the sample solution! 11. The reaction vessel (510) according to claim 10, wherein the reaction substrate (511) extends substantially parallel to the flow direction of the sample solution.  [12] It has a flow path member (512) that cooperates with the reaction substrate (511) to define the flow path, and the flow path member (512) includes two through holes (512a, 512b) and the two A groove (512c) extending between the two through holes (512a, 512b), and the elastic member (514) seals one through hole (512b) of the flow path member (512). 12. The reaction vessel (510) according to claim 11, wherein the reaction vessel (510) is stopped.  [13] A reactor (120; 220) using the reaction vessel (110; 210; 410; 510) according to claim 1.  ; 420; 520),  A reaction vessel housing part (130; 430; 530) for housing the reaction vessel (110; 210; 410; 510);  And a drive unit (140; 240; 440; 540) for deforming the elastic member (114; 214; 514) of the reaction vessel (110; 210; 410; 510). Reactor (120; 220; 420; 520).  [14] The reaction device (120; 220) according to claim 13, wherein the driving unit (140; 240; 540) deforms the elastic member (114; 214; 514) by mechanical means. 520).  15. The reaction device (420) according to claim 13, wherein the drive unit (440) deforms the elastic member (214) by fluid means.