Reactor plate and reaction processing method

Disclosed herein is a reactor plate which prevents the entry of foreign matter from outside and the pollution of an outside environment. The reactor plate includes a sealed reaction well (5), reaction well channels (13, 15, 17) connected to the reaction well (5), and a syringe (51) for sending a liquid to the reaction well channels (13, 15, 17) and the reaction well (5). The syringe (51) has a cylinder (51a), a plunger (51b), and a cover body (51d). The cover body (51d) has flexibility in the sliding direction of the plunger (51b), and is connected to the cylinder (51a) and the plunger (51b) to create a sealed space (51e) enclosed with the cylinder (51a), the plunger (51b), and the cover body (51d). The cover body (51d) is provided to hermetically cut off a part of an inner wall of the cylinder (51a) to be brought into contact with the plunger (51b) from an atmosphere outside the cylinder (51a).

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to a reactor plate suitable for use in various assays and analyses such as biological and biochemical assays and general chemical analyses in the fields of medical care and chemistry, and a reaction processing method for processing such a reactor plate.

2. Description of the Related Art

As small reactors for use in biochemical assays or general chemical analyses, micro multi-chamber devices are used. Examples of such devices include micro well reactor plates such as a microtiter plate constituted from a plate-shaped substrate having a plurality of wells formed in the surface thereof (see, for example, Patent Document 1) and the like.

Further, as a structure for dispensing a small amount of liquid which can quantitatively treat a small amount of liquid, a structure having a first channel, a second channel, a third channel which is in communication with the first channel through an opening provided in the channel wall of the first channel, and a fourth channel which is in communication with the second channel through an opening provided in the channel wall of the second channel, connects one end of the third channel to the second channel, and has relatively lower capillary attraction than the third channel is developed (see, for example, Patent Documents 2, 3). When such a structure for dispensing a small amount of liquid is used, a liquid introduced into the first channel is drawn into the third channel, and then the liquid remaining in the first channel is removed, and as a result, the liquid having a volume corresponding to the capacity of the third channel is dispensed into the second channel.Patent Document 1: Japanese Patent Application Laid-open No. 2005-177749Patent Document 2: Japanese Patent Application Laid-open No. 2004-163104Patent Document 3: Japanese Patent Application Laid-open No. 2005-114430Patent Document 3: Japanese Patent No. 3452717

DISCLOSURE OF THE INVENTION

Problem to be Solved by the Invention

When a conventional micro well reactor plate is used, the top surface of the reactor plate is open to the atmosphere. Therefore, there is a fear that foreign matter will enter a sample from outside, or on the other hand, a reaction product will pollute an outside environment.

Further, in the structure for dispensing a small amount of liquid disclosed in Patent Documents 2 and 3, each of the first and second channels has a port for introducing a liquid at each end thereof. However, these ports are open to the atmosphere and, therefore, there is a possibility that a reaction product will leak through the ports and then pollute an outside environment.

Therefore, it is an object of the present invention to provide a reactor plate which can prevent the entry of foreign matter from outside and the pollution of an outside environment, and a reaction processing method using such a reactor plate.

Means for Solving the Problem

The present invention is directed to a reactor plate including a sealed reaction well, a reaction well channel connected to the reaction well, and a syringe for sending a liquid to the reaction well channel and the reaction well. The syringe has a cylinder having a discharge port connected to the reaction well channel, a plunger placed in the cylinder, and a cover body for hermetically cutting off a part of an inner wall of the cylinder to be brought into contact with the plunger from an atmosphere outside the cylinder. The cover body has flexibility in the sliding direction of the plunger, and is connected to the cylinder and the plunger to create a sealed space enclosed with the cylinder, the plunger, and the cover body.

In the reactor plate according to the present invention, since the reaction well is sealed and an internal space of the cylinder connected to the reaction well through the discharge port of the cylinder and the reaction well channel is sealed with the plunger, it is possible to prevent the entry of foreign matter from the outside of the reactor plate and the pollution of an outside environment with a liquid.

Further, since the syringe has the cover body for hermetically cutting off a part of an inner wall of the cylinder to be brought into contact with the plunger from an atmosphere outside the cylinder, it is possible to prevent the entry of foreign matter from outside through a gap between the cylinder and the plunger. In addition, it is also possible to prevent a liquid from leaking out of the reactor plate through the gap between the cylinder and the plunger, thereby preventing the pollution of an outside environment with the liquid.

Here, the cover body needs to have flexibility in at least the sliding direction of the plunger, but may have flexibility also in a direction other than the sliding direction of the plunger.

The reactor plate according to the present invention may further include a variable capacity part whose internal space is sealed and whose internal capacity is passively variable and a syringe air vent channel whose one end is connected to the sealed space and whose other end is connected to the variable capacity part.

The reactor plate according to the present invention may further include a sealed well provided separately from the reaction well, a sealed well channel connected to the sealed well, and a switching valve for connecting the syringe to the reaction well channel or the sealed well channel.

An example of the sealed well includes a sample well for containing a sample liquid.

For example, the sample well may be hermetically sealed with an elastic member which allows a dispensing device having a sharp tip to pass through to form a through hole and which also allows the through hole to be closed by pulling out the dispensing device due to its elasticity.

Further, the sample well may previously contain a liquid for pretreating a sample or a reagent.

The reactor plate according to the present invention may further include one or more reagent wells, each of which is constituted from the sealed well, other than the sample well. The reagent well previously contains a reagent to be used for the reaction of a sample liquid and is sealed with a film, or has an openable and closable cap so that the reagent can be injected thereinto. An example of the film for sealing the reagent well to prevent the leakage of the reagent includes one through which a dispensing device having a sharp tip can pass.

In a case where the reactor plate according to the present invention is intended to be used for gene analysis, the reactor plate preferably includes a gene amplification well which is constituted from the sealed well and used for carrying out gene amplification reaction. The gene amplification well preferably has a shape suitable for controlling a temperature according to a predetermined temperature cycle. It is to be noted that gene amplification can also be carried out in the reaction well.

An example of the switching valve includes a rotary valve. The rotary valve may have a port to be connected to the syringe at the center of rotation. In this case, the syringe may be placed on the rotary valve.

The reactor plate according to the present invention may further include a reaction well air vent channel connected to the reaction well. As a specific example of the channel configuration of the reactor plate according to the present invention, the reaction well channel which is constituted from a groove formed in the contact surface between two members bonded together, or from the groove and a through hole formed in both or one of the members and which includes a main channel connected to the syringe, a metering channel branched off the main channel and having a predetermined capacity, and an injection channel whose one end is connected to the metering channel and whose other end is connected to the reaction well can be mentioned. In this case, the main channel and the reaction well air vent channel can be hermetically sealed, and the injection channel is formed narrower than the metering channel, and does not allow the passage of a liquid at a liquid introduction pressure applied to introduce the liquid into the main channel and the metering channel and at a purge pressure applied to purge the liquid from the main channel but allows the passage of the liquid at a pressure higher than the liquid introduction pressure and the purge pressure.

Here, the phrase “the injection channel is formed narrower than the metering channel” means that in a case where the injection channel is constituted from a plurality of channels, each of the channels constituting the injection channel is formed narrower than the metering channel.

The present invention is also directed to a reaction processing method using the reactor plate according to the present invention having the channel configuration described above as an example of a channel configuration, the method including filling the main channel and the metering channel with a liquid at the introduction pressure, purging the liquid from the main channel by flowing a gas through the main channel while allowing the liquid to remain in the metering channel, and injecting the liquid contained in the metering channel into the reaction well through the injection channel by creating a positive pressure higher than the introduction pressure in the main channel, or by creating a negative pressure in the reaction well, or by creating a positive pressure higher than the introduction pressure in the main channel and creating a negative pressure in the reaction well.

In the channel configuration described above as an example of a channel configuration, the contact angle of the injection channel with a water drop is, for example, 90° or larger, and the area of an interface between the injection channel and the metering channel is, for example, 1 to 10,000,000 μm2(square micrometers). It is to be noted that in a case where the injection channel is constituted from a plurality of channels, the phrase “the area of an interface between the injection channel and the metering channel” means the area of an interface between each of the channels constituting the injection channel and the metering channel.

The reactor plate according to the present invention may include a plurality of the reaction wells. In this case, the metering channel and the injection channel may be provided for each of the reaction wells, and the plurality of metering channels may be connected to the main channel.

Further, a projecting portion may be provided so as to project from a top inner surface of the reaction well. In this case, the other end of the injection channel is located at the tip of the projecting portion. The projecting portion includes one having a proximal end and a distal end narrower than the proximal end.

The reaction well can be used for carrying out at least any one of color reaction, enzymatic reaction, fluorescence reaction, chemiluminescence reaction, and bioluminescence reaction.

In a case where the reactor plate according to the present invention is intended to be used for measuring a gene-containing sample, a sample previously subjected to gene amplification reaction may be introduced into the reactor plate, or a gene amplification reagent may be previously contained in the reaction well or the reactor plate may be designed to allow a gene amplification reagent to be dispensed into the reaction well so that gene amplification reaction can be carried out in the reaction well of the reactor plate.

Examples of the gene amplification reaction include PCR method and LAMP method. As PCR method for amplifying DNA, a method is proposed for directly subjecting a sample such as blood to PCR reaction without pretreating the sample. More specifically, this method is a nucleic acid synthesis method for amplifying a target gene contained in a gene-containing sample by adding a gene-containing body contained in the gene-containing sample or the gene-containing sample itself and then adjusting the pH of the thus obtained reaction mixture to 8.5 to 9.5 (25° C.) (see Patent Document 4).

The reaction well may be made of an optically-transparent material so that optical measurement can be carried out from the bottom of the reaction well or from above the reaction well.

In a case where a liquid to be introduced into the reaction well channel contains a gene, the reaction well may contain a probe which reacts with the gene.

Further, the probe may be fluorescently-labeled.

Effect of the Invention

As described above, the reactor plate according to the present invention includes a sealed reaction well, a reaction well channel connected to the reaction well, and a syringe for sending a liquid to the reaction well channel and the reaction well, the reaction well is sealed, and an internal space of the cylinder connected to the reaction well through the discharge port of the cylinder and the reaction well channel is sealed with the plunger. Therefore, it is possible to prevent the entry of foreign matter from the outside of the reactor plate and the pollution of an outside environment with a liquid.

Further, the syringe has a cylinder having a discharge port connected to the reaction well channel, a plunger placed in the cylinder, and a cover body for hermetically cutting off a part of an inner wall of the cylinder to be brought into contact with the plunger from an atmosphere outside the cylinder, and the cover body has flexibility in the sliding direction of the plunger and is connected to the cylinder and the plunger to create a sealed space enclosed with the cylinder, the plunger, and the cover body. Therefore, it is possible to prevent the entry of foreign matter from outside through a gap between the cylinder and the plunger. In addition, it is also possible to prevent a liquid from leaking out of the reactor plate through the gap between the cylinder and the plunger, thereby preventing the pollution of an outside environment with the liquid.

In a case where the reactor plate according to the present invention is intended to be used for measuring a gene-containing sample, the sample injected into the reactor plate and then introduced into the reaction well can be processed in a closed system. Therefore, it is possible to prevent the pollution of an environment outside the reactor plate and the contamination of the sample with foreign matter coming from outside the reactor plate.

The reactor plate according to the present invention may further include a variable capacity part whose internal space is sealed and whose internal capacity is passively variable and a syringe air vent channel whose one end is connected to the sealed space and whose other end is connected to the variable capacity part. This makes it possible to cut off the sealed space from an atmosphere outside the reactor plate and to relieve a pressure change in the sealed space caused by a change in the internal capacity of the sealed space during the sliding of the plunger, thereby making it possible to smoothly slide the plunger.

The reactor plate according to the present invention may further include a sealed well provided separately from the reaction well, a sealed well channel connected to the sealed well, and a switching valve for connecting the syringe to the reaction well channel or the sealed well channel.

For example, by providing a sample well for containing a sample liquid as the sealed well, it is possible to eliminate the necessity to separately prepare a well for containing a sample.

Further, the sample well may be hermetically sealed with an elastic member which allows a dispensing device having a sharp tip to pass through to form a through hole and which also allows the through hole to be closed by pulling out the dispensing device due to its elasticity. This makes it possible to inject a sample liquid into the sample well sealed with the elastic member and to prevent the sample liquid injected into the sample well from leaking out of the sample well.

Further, the sample well may previously contain a liquid for pretreating a sample or a reagent. This makes it possible to eliminate the necessity to dispense a liquid for pretreating a sample or a reagent into the sample well.

The reactor plate according to the present invention may further include one or more reagent wells, each of which is constituted from the sealed well, other than the sample well. By allowing the reagent well to previously contain a reagent to be used for the reaction of a sample liquid and sealing it with a film or by allowing the reagent well to have an openable and closable cap so that the reagent can be injected thereinto, it is possible to eliminate the necessity to separately prepare a well for containing the reagent.

The reactor plate according to the present invention may further include a gene amplification well which is constituted from the sealed well and used for carrying out gene amplification reaction. By providing such a gene amplification well, it is possible to amplify a target gene in the reactor plate by gene amplification reaction such as PCR method or LAMP method even when a sample liquid contains only a very small amount of the target gene, thereby increasing analytical precision.

The reactor plate according to the present invention may further include a sealed well channel connected to the sealed well, a syringe for sending a liquid, and a switching valve for connecting the syringe to the main channel or the sealed well channel. In this case, a liquid contained in the sealed well can be injected into the main channel with the syringe and the switching valve.

The switching valve may be a rotary valve. In this case, by providing a port to be connected to the syringe at the center of rotation of the rotary valve, it is possible to simplify a channel configuration.

Further, by providing a port to be connected to the syringe at the center of rotation of the rotary valve and placing the syringe on the rotary valve, it is possible to shorten or eliminate a channel between the port and the syringe, thereby simplifying the structure of the reactor plate. In addition, it is also possible to effectively utilize a region on the switching valve, thereby making it possible to make the planar size of the reactor plate smaller as compared to a case where the syringe is placed in a region other than the region on the switching valve.

The reactor plate according to the present invention may further include a reaction well air vent channel connected to the reaction well. As a specific example of the channel configuration of the reactor plate according to the present invention, the reaction well channel which is constituted from a groove formed in the contact surface between two members bonded together, or from the groove and a through hole formed in both or one of the members and which includes a main channel connected to the syringe, a metering channel branched off the main channel and having a predetermined capacity, and an injection channel whose one end is connected to the metering channel and whose other end is connected to the reaction well can be mentioned. In this case, the main channel and the reaction well air vent channel can be hermetically sealed, and the injection channel is formed narrower than the metering channel, and does not allow the passage of a liquid at a liquid introduction pressure applied to introduce the liquid into the main channel and the metering channel and at a purge pressure applied to purge the liquid from the main channel but allows the passage of the liquid at a pressure higher than the liquid introduction pressure and the purge pressure. By carrying out the reaction processing method according to the present invention with the reactor plate according to the present invention, it is possible to prevent the entry of foreign matter from the outside of the reactor plate and the pollution of an outside environment with a liquid.

Further, by providing the reaction well air vent channel connected to the reaction well, it is possible to move a gas between the reaction well and the reaction well air vent channel when a liquid is injected into the reaction well through the injection channel, thereby making it possible to smoothly inject a liquid into the reaction well. The reaction well air vent channel can also be used to suck a gas contained in the reaction well to decompress the reaction well to inject a liquid into the reaction well.

In the channel configuration described above as an example of a channel configuration, the contact angle of the metering channel and the injection channel with a water droplet is preferably 90° or larger, and the area of an interface between the injection channel and the metering channel is preferably 1 to 10,000,000 μm2. This makes it difficult for a liquid to enter the injection channel when the liquid is introduced into the main channel and the metering channel, thereby making it possible to increase an introduction pressure applied to introduce a liquid into the main channel and the metering channel.

The reactor plate according to the present invention may include a plurality of the reaction wells. In this case, by providing the metering channel and the injection channel for each of the reaction wells and connecting the plurality of metering channels to the main channel, it is possible to introduce a liquid into the plurality of metering channels one after another and then simultaneously inject the liquid into the plurality of reaction wells through the injection channels.

Further, a projecting portion may be provided so as to project from a top inner surface of the reaction well. In this case, the other end of the injection channel is located at the tip of the projecting portion. By allowing the projecting portion to have a proximal end and a distal end narrower than the proximal end, a liquid to be injected into the reaction well through the injection channel can be easily dropped into the reaction well.

In a case where the reactor plate according to the present invention is intended to be used for measuring a gene-containing sample, the reactor plate may be designed to allow gene amplification reaction to be carried out in the reaction well. This makes it possible to eliminate the necessity to prepare a sample which has been subjected to gene amplification reaction outside the reactor plate.

The reaction well may be made of an optically-transparent material so that optical measurement can be carried out from the bottom of the reaction well or from above the reaction well. This makes it possible to optically measure a liquid contained in the reaction well without transferring the liquid into another well.

In a case where a liquid to be introduced into the reaction well channel contains a gene, the reaction well may contain a probe which reacts with the gene. This makes it possible to detect a gene having a base sequence corresponding to the probe in the reaction well.

DESCRIPTION OF THE REFERENCE NUMERALS

DETAILED DESCRIPTION OF THE INVENTION

FIG. 1Ais a schematic plan view of one embodiment of a reactor plate according to the present invention, andFIG. 1Bis a schematic sectional view taken along the A-A line inFIG. 1A, which further includes the sectional views of a metering channel15, an injection channel17, reaction well air vent channels19and21, a liquid drain space29, an air drain space31, and a bellows53.FIG. 1Cis a schematic expanded sectional view of a syringe51and the bellows53of the reactor plate in the embodiment shown inFIG. 1Aand their vicinity.FIG. 2shows an exploded sectional view of the reactor plate in the embodiment shown inFIG. 1Aand a schematic exploded perspective view of a switching valve.FIGS. 3A to 3Care a schematic plan view, a schematic perspective view, and a schematic sectional view of one reaction well of the reactor plate in the embodiment shown inFIG. 1Aand its vicinity, respectively.FIG. 4Ais an expanded plan view of a sample well, andFIG. 4Bis a sectional view taken along the B-B line inFIG. 4A.FIG. 5Ais an expanded plan view of a reagent well, andFIG. 5Bis a sectional view taken along the C-C line inFIG. 5A.FIG. 6Ais an expanded plan view of a well for air suction, andFIG. 6Bis a sectional view taken along the D-D line inFIG. 6A. With reference toFIGS. 1A to 6B, the reactor plate according to one embodiment of the present invention will be described.

A reactor plate1includes a plurality of reaction wells5each having an opening in one surface of a well base3. In the reactor plate1according to this embodiment of the present invention, the reaction wells5are arranged in an array of 6 rows and 6 columns in a staggered format. In each of the reaction wells5, a reagent7and a wax9are contained.

The material of the well base3including the reaction wells5is not particularly limited. However, in a case where the reactor plate1is intended to be disposable, the material of the well base3is preferably a cheaply-available material. Preferred examples of such a material include resin materials such as polypropylene and polycarbonate. In a case where the reactor plate1is intended to be used to detect a substance in the reaction well5by absorbance, fluorescence, chemiluminescence, or bioluminescence, the container base3is preferably made of an optically-transparent resin so that optical detection can be carried out from the bottom of the reaction well5. Particularly, in a case where the reactor plate1is intended to be used for fluorescence detection, the container base3is preferably made of a low self-fluorescent (i.e., fluorescence emitted from a material itself is weak) and optically-transparent resin, such as polycarbonate. The thickness of the well base3is in a range of 0.2 to 4.0 mm (millimeters), preferably in a range of 1.0 to 2.0 mm. From the viewpoint of low self-fluorescence, the thickness of the well base3for fluorescence detection is preferably small.

Referring toFIGS. 1A,1B,1C,3A,3B and3C, a channel base11is provided on the well base3so as to cover a region where the reaction wells5are arranged. The channel base11is made of, for example, PDMS (polydimethylsiloxane) or silicone rubber. The thickness of the channel base11is, for example, from 1.0 to 5.0 mm. The channel base11has a groove in its surface which is in contact with the well base3. The groove and the surface of the well base3together form a main channel13, the metering channel15, the injection channel17, the reaction well air vent channels19and21, and drain space air vent channels23and25. The main channel13, the metering channel15, and the injection channel17constitute a reaction well channel. In the surface of the channel base11which is in contact with the well base3, a recess27is also provided so as to be located above each of the reaction wells5. It is noted that, inFIG. 1AandFIGS. 3A and 3B, the channel base11is not shown, and only the groove and recess provided in the channel base11are shown.

The main channel13is constituted from one channel, and is therefore bent so as to run in the vicinity of all the reaction wells5. One end of the main channel13is connected to a channel13aconstituted from a through hole provided in the well base3. The channel13ais connected to a port of a switching valve63(which will be described later). The other end of the main channel13is connected to the liquid drain space29provided in the well base3. The main channel13is constituted from a groove having a depth of, for example, 400 μm (micrometers) and a width of, for example, 500 μm. It is noted that a part of the main channel13having a predetermined length (e.g., 250 μm) and located downstream of a position, to which the metering channel15is connected, has a width smaller than that of the other part of the main channel13, and the width of such a part is, for example, 250 μm.

The metering channel15branches off the main channel13, and is provided for each of the reaction wells5. The end of the metering channel15on the opposite side from the main channel13is located in the vicinity of the reaction well5. The depth of a groove constituting the metering channel15is, for example, 400 μm. The metering channel15is designed to have a predetermined internal capacity of, for example, 2.5 μL (microliters). A part of the metering channel15connected to the main channel13has a width larger than that of the above-described narrow part of the main channel13(e.g., 500 μm). Therefore, at a position where the metering channel15branches off the main channel13, the resistance to the flow of a liquid coming from one end of the main channel13is larger in the main channel13than in the metering channel15. For this reason, the liquid coming from one end of the main channel13first flows into the metering channel15to fill the metering channel15, and then flows downstream through the narrow part of the main channel13.

The injection channel17is also provided for each of the reaction wells5. One end of the injection channel17is connected to the metering channel15, and the other end of the injection channel17is connected to the recess27located above the reaction well5so as to be led to the space above the reaction well5. The injection channel17is designed to have a size allowing the liquid-tightness of the reaction well5to be maintained in a state where there is no difference between the pressure in the reaction well5and the pressure in the injection channel17. According to the present embodiment, the injection channel17is constituted from a plurality of grooves, and each groove has a depth of, for example, 10 μm and a width of, for example, 20 μm, and the pitch between the adjacent grooves is, for example, 20 μm, and the thirteen grooves are provided in a region having a width of 500 μm. In this case, the area of an interface between the groove constituting the injection channel17and the metering channel15, that is, the cross-sectional area of the groove constituting the injection channel17is 200 μm2. The recess27has a depth of, for example, 400 μm, and has a circular planar shape smaller than that of the reaction well5.

The reaction well air vent channel19is provided for each of the reaction wells5. One end of the reaction well air vent channel19is connected to the recess27, which is located above the reaction well5, at a position different from the position, to which the injection channel17is connected, so as to be located above the reaction well5. The reaction well air vent channel19is designed to have a size allowing the liquid-tightness of the reaction well5to be maintained in a state where there is no difference between the pressure in the reaction well5and the pressure in the reaction well air vent channel19. The other end of the reaction well air vent channel19is connected to the reaction well air vent channel21. According to the present embodiment, the reaction well air vent channel19is constituted from a plurality of grooves, and each groove has a depth of, for example, 10 μm and a width of, for example, 20 μm, and the pitch between the adjacent grooves is, for example, 20 μm, and the thirteen grooves are provided in a region having a width of 500 μm.

The reactor plate according to the present embodiment has plurality a of the reaction well air vent channels21. To each of the reaction well air vent channels21, the plurality of reaction well air vent channels19are connected. These reaction well air vent channels21are provided to connect the reaction well air vent channels19to the air drain space31provided in the well base3. Each of the reaction well air vent channels21is constituted from a groove having a depth of, for example, 400 μm and a width of, for example, 500 μm.

The drain space air vent channel23is provided to connect the liquid drain space29to a port of the switching valve63(which will be described later). One end of the drain space air vent channel23is located above the liquid drain space29. The other end of the drain space air vent channel23is connected to a channel23aconstituted from a through hole provided in the well base3. The channel23ais connected to a port of the switching valve63(which will be described later). The drain space air vent channel23is constituted from a groove having a depth of, for example, 400 μm and a width of, for example, 500 μm.

The drain space air vent channel25is provided to connect the air drain space31to a port of the switching valve63(which will be described later). One end of the drain space air vent channel25is located above the air drain space31. The other end of the drain space air vent channel25is connected to a channel25aconstituted from a through hole provided in the well base3. The channel25ais connected to a port of the switching valve63(which will be described later). The drain space air vent channel25is constituted from a groove having a depth of, for example, 400 μm and a width of, for example, 500 μm.

On the channel base11, a channel cover33(not shown inFIG. 1A) is provided. The channel cover33is provided to fix the channel base11to the well base3. The channel cover33has a through hole formed to be located above each of the reaction wells5.

Referring toFIGS. 1A,1B,1C,4A and4B, in the well base3, a sample well35, a reagent well37, and a well39for air suction are provided at positions other than the positions of a region where the reaction wells5are arranged, and the drain spaces29and31. The sample well35, the reagent well37, and the well39for air suction constitute sealed wells of the reactor plate according to the present invention.

In the well base3, a sample channel35aconstituted from a through hole extending from the bottom of the sample well35to the back surface of the well base3and a sample well air vent channel35bconstituted from a through hole extending from the top surface to the back surface of the well base3are provided in the vicinity of the sample well35. On the well base3, a projecting portion35cis provided so as to surround an opening of the sample well35. In the projecting portion35c, a sample well air vent channel35dconstituted from a through hole is provided so as to be located above the sample well air vent channel35b. In the surface of the projecting portion35c, a sample well air vent channel35ewhich allows the sample well35to communicate with the sample well air vent channel35dis provided.

The sample well air vent channel35eis constituted from one or more narrow holes, and each narrow hole has a width of, for example, 5 to 200 μm and a depth of, for example, 5 to 200 μm. The sample well air vent channel35eis provided to maintain the liquid-tightness of the sample well35in a state where there is no difference between the pressure in the sample well35and the pressure in the sample well air vent channel35d. On the projecting portion35c, a septum41as an elastic member to cover the sample well35and the air vent channel35dis provided. The septum41is made of an elastic material such as silicone rubber or PDMS. Therefore, a dispensing device having a sharp tip can pass through the septum41to form a through hole, but the through hole can be closed by pulling the dispensing device out of the septum41due to its elasticity. On the septum41, a septum stopper43for fixing the septum41is provided. The septum stopper43has an opening located above the sample well35. According to the present embodiment, a reagent45is previously contained in the sample well35.

As shown inFIGS. 5A and 5B, in the well base3, a reagent channel37aconstituted from a through hole extending from the bottom of the reagent well37to the back surface of the well base3and a reagent well air vent channel37bconstituted from a through hole extending from the top surface to the back surface of the well base3are provided in the vicinity of the reagent well37. On the well base3, a projecting portion37cis provided so as to surround an opening of the reagent well37. In the projecting portion37c, a reagent well air vent channel37dconstituted from a through hole is provided so as to be located above the reagent well air vent channel37b. In the surface of the projecting portion37c, a reagent well air vent channel37ewhich allows the reagent well37to communicate with the reagent well air vent channel37dis provided.

The reagent well air vent channel37eis constituted from one or more narrow holes, and each narrow hole has a width of, for example, 5 to 200 μm and a depth of, for example, 5 to 200 μm. The reagent well air vent channel37eis provided to maintain the liquid-tightness of the reagent well37in a state where there is no difference between the pressure in the reagent well37and the pressure in the reagent well air vent channel37d. On the projecting portion37c, a film47made of, for example, aluminum to cover the reagent well37and the air vent channel37dis provided. In the reagent well37, dilution water49is contained.

As shown inFIGS. 6A and 6B, the well39for air suction has the same structure as the reagent well37. That is, in the well base3, a channel39afor air suction constituted from a through hole extending from the bottom of the well39for air suction to the back surface of the well base3and an air vent channel39bfor the well for air suction constituted from a through hole extending from the top surface to the back surface of the well base3are provided in the vicinity of the well39for air suction. On the well base3, a projecting portion39chaving air vent channels39dand39efor the well for air suction is provided so as to surround an opening of the well39for air suction. On the projecting portion39c, a film47made of, for example, aluminum is provided. The well39for air suction contains neither a liquid nor a solid, but is filled with air.

Referring toFIGS. 1A,1B,1C, and2, a syringe51is provided in the surface of the well base3at a position other than the positions of a region where the reaction wells5are arranged, the drain spaces29and31, and the wells35,37and39. The syringe51is constituted from a cylinder51aprovided in the well base3and having a discharge port provided at the bottom thereof, a plunger51bplaced in the cylinder51a, and a cover body51d. In the well base3, a syringe channel51cextending from the discharge port of the cylinder51ato the back surface of the well base3is provided.

The cover body51dhas flexibility in the sliding direction of the plunger51b, and is connected to the cylinder51aand the plunger51b. The cover body51dis provided to create a sealed space51eto hermetically cut off a part of an inner wall of the cylinder51ato be brought into contact with the plunger51bfrom an atmosphere outside the cylinder51a. The sealed space51eis enclosed with the cylinder51a, the plunger51b, and the cover body51d. One end of the cover body51dconnected to the cylinder51ais hermetically fixed to the upper end of the cylinder51aby a cylinder cap51f. On the other hand, the other end of the cover body51dconnected to the plunger51bis hermetically connected to the upper surface of the plunger51bby an adhesive. However, a method for connecting the cover body51dto the cylinder51aand the plunger51bis not limited to the method described above, and the connecting positions of the cover body51dare not limited to those described above.

As described above, the cover body51dis connected to the cylinder51aand the plunger51bto create a sealed space51eenclosed with the cylinder51a, the plunger51b, and the cover body51dand, therefore, it is possible to prevent the entry of foreign matter from outside through a gap between the cylinder51aand the plunger51b. In addition, it is also possible to prevent the leakage of a liquid through the gap between the cylinder51aand the plunger51b, thereby preventing the pollution of an outside environment with the liquid. It is to be noted that as described above, since the cover body51dhas flexibility in the sliding direction of the plunger51b, the plunger51bcan be slidably moved.

According to the present embodiment, the plunger51band the cover body51dare provided as separate members, but the plunger and the cover body may be integrally molded. The plunger and the cover body can be integrally molded using, for example, silicone rubber.

In the well base3, the bellows53is also provided at a position other than the positions of a region where the reaction wells5are arranged, the drain spaces29and31, the wells35,37, and39, and the syringe51. The bellows53has a sealed internal space, and the internal capacity of the bellows53is passively variable by extraction and contraction. The bellows53is placed in, for example, a through hole53aprovided in the well base3.

A well bottom55is attached to the back surface of the well base3at a position other than the position of a region where the reaction wells5are arranged. In the well bottom55, an air vent channel53bis provided at a position allowing the air vent channel53bto communicate with the bellows53. The bellows53is connected to the well bottom55so as to be in close contact with the surface of the well bottom55. The well bottom55is provided to guide the channels13a,23a,25a,35a,35b,37a,37b,39a,39b,51c, and53bto predetermined port positions.

In the well base3and the well bottom55, a syringe air vent channel53cwhose one end is connected to the sealed space51eand whose other end is connected to the bellows53is provided. InFIG. 1A, the syringe air vent channel53cis not shown.

As described above, the reactor plate1has the syringe air vent channel53cwhose one end is connected to the sealed space51eand whose other end is connected to the bellows53and, therefore, it is possible to cut off the sealed space51efrom an atmosphere outside the reactor plate1and to relieve a pressure change in the sealed space51ecaused by a change in the internal capacity of the sealed space51eduring the sliding of the plunger51b, thereby making it possible to smoothly slide the plunger51b.

On the surface of the reaction well bottom55located on the opposite side from the well base3, the rotary switching valve63is provided. The switching valve63is constituted from disk-shaped sealing plate57, rotor upper59, and rotor base61. The switching valve63is attached to the well bottom55by means of a lock65.

The sealing plate57has a through hole57a, a through groove57b, and a through hole57c. The through hole57ais provided in the vicinity of the peripheral portion of the sealing plate57, and is connected to any one of the channels13a,35a,37a, and39a. The through groove57bis provided inside the through hole57aand on a circle concentric with the sealing plate57, and is connected to at least two of the channels23a,25a,35b,37b,39b, and53b. The through hole57cis provided at the center of the sealing plate57, and is connected to the syringe channel51c.

The rotor upper59has a through hole59a, a groove59h, and a through hole59c. The through hole59ais provided at a position corresponding to the through hole57aprovided in the sealing plate57. The groove59bis provided in the surface of the rotor upper59so as to correspond to the through groove57bprovided in the sealing plate57. The through hole59cis provided at the center of the rotor upper59.

The rotor base61has a groove61a. The groove61ais provided in the surface of the rotor base61to connect the through hole59aprovided in the peripheral portion of the rotor upper59and the through hole59cprovided at the center of the rotor upper59to each other.

By rotating the switching valve63, the syringe channel51cis connected to any one of the channels13a,35a,37a, and39a, and at the same time, the air vent channel53bis also connected to at least any one of the channels23a,25a,35b,37b, and39b.

The switching valve63shown inFIG. 1Ais in its initial state where the syringe channel51cis not connected to any one of the channels13a,35a,37a, and39a, and the air vent channel53bis not connected to any one of the channels23a,25a,35b,37b, and39b, either.

As described above, the injection channel17provided in the reactor plate1is designed so that the liquid-tightness of the reaction well5is maintained in a state where there is no difference between the pressure in the injection channel17and the pressure in the reaction well5. The reaction well air vent channel19is also designed so that the liquid-tightness of the reaction well5is maintained in a state where there is no difference between the pressure in the reaction well5and the pressure in the reaction well air vent channel19. The main channel13constituting the reaction well channel, the liquid drain space29connected to the main channel13, and the drain space air vent channel23can be hermetically sealed by switching of the switching valve63. The wells35,37, and39are sealed with the septum41or the film47. The channels35a,35b,37a,37b,39a, and39bconnected to the wells35,37, and39, respectively, can be hermetically sealed by switching the switching valve63. One end of the air vent channel53bis connected to the bellows53and, therefore, the air vent channel53bis hermetically sealed. As described above, the wells and channels in the reactor plate1constitute a closed system. It is noted that even in a case where the reactor plate1does not have the bellows53and the air vent channel53bis connected to the atmosphere outside the reactor plate1, the air vent channel53bcan be cut off from the wells and the channels other than the air vent channel53bprovided in the reactor plate1by switching of the switching valve63and, therefore, the wells for containing a liquid and the channels for flowing a liquid can be hermetically sealed.

FIG. 7is a sectional view showing a reaction processing apparatus for processing the reactor plate1shown inFIGS. 1A,1B and1C as well as the reactor plate1. The reactor plate1shown inFIG. 7has the same structure as that shown inFIGS. 1A,1B and1C and, therefore, the description thereof is omitted.

The reaction processing apparatus includes a temperature control system for controlling the temperature of the reaction wells5, a syringe driving unit69for driving the syringe51, and a switching valve driving unit71for switching the switching valve63.

FIGS. 8 to 14are plan views for explaining the operation of introducing a sample liquid into the reaction wells5from the sample well35. This operation will be described with reference to FIGS.1A-1C and8to14.

A dispensing device having a sharp tip (not shown) is prepared, and the dispensing device is passed through the septum41provided on the sample well35to dispense, for example, 5 μL of a sample liquid into the sample well35. After the completion of the dispensing of the sample liquid, the dispensing device is pulled out of the septum41. By pulling the dispensing device out of the septum41, a through hole formed in the septum41is closed due to the elasticity of the septum41.

The syringe driving unit69is connected to the plunger51bof the syringe51, and the switching valve driving unit71is connected to the switching valve63.

As shown inFIG. 8, the switching valve63in its initial state shown inFIG. 1Ais rotated to connect the syringe channel51cto the sample channel35aand to connect the air vent channel53bto the sample well air vent channel35b. At this time, the air vent channels37band39bare also connected to the air vent channel53b. The sample well35contains, for example, 45 μL of a reagent45.

The plunger51bof the syringe51is allowed to slide to mix the sample liquid and the reagent45contained in the sample well35. Then, for example, only 10 μL of the liquid mixture contained in the sample well35is sucked into the channel in the switching valve63, the syringe channel51c, and the syringe51. At this time, the bellows53expands and contracts with changes in the volume of a gas contained in the sample well35because the sample well35is connected the bellows53through the air vent channels35e,35d, and35b, the switching valve63, and the air vent channel53b. Further, the cover body51dis deformed by sliding the plunger51bso that the internal capacity of the sealed space51e(seeFIG. 1C) is changed. The bellows53expands and contracts also with changes in the internal capacity of the sealed space51ebecause the sealed space51eis connected to the bellows53through the syringe air vent channel53c. Also in the operation steps which will be described below, the bellows53expands and contracts with changes in the internal capacity of the sealed space51edue to the sliding of the plunger51b.

As shown inFIG. 9, the switching valve63is rotated to connect the syringe channel51cto the reagent channel37aand to connect the air vent channel53bto the reagent well air vent channel37b. The reagent well37contains, for example, 190 μL of dilution water49. The mixture sucked into the channel in the switching valve63, the syringe channel51c, and the syringe51is injected into the reagent well37. Then, the syringe51is slidably moved to mix the mixture and the dilution water49. For example, the whole diluted mixture, that is, 200 μL of the diluted mixture is sucked into the channel in the switching valve63, the syringe channel51c, and the syringe51. At this time, the bellows53expands and contracts with changes in the volume of a gas contained in the reagent well37, since the reagent well37is connected to the bellows53through the air drain channels37e,37d, and37b, the switching valve63, and the air vent channel53b.

As shown inFIG. 10, the switching valve63is rotated to connect the syringe channel51cto the channel13aconnected to one end of the main channel13and to connect the air vent channel53bto the channels23aand25aconnected to the liquid drain space29and the air drain space31, respectively. The syringe51is driven in an extrusion direction to send the diluted mixture sucked into the channel in the switching valve63, the syringe channel51c, and the syringe51to the main channel13. As shown by the arrows and dots inFIG. 10, the diluted mixture injected into the main channel13through the channel13afills the metering channels15one after another in order of increasing distance from the channel13a, and then reaches the liquid drain space29. The injection channel17allows the passage of a gas but does not allow the passage of the diluted mixture at an introduction pressure applied to introduce the diluted mixture into the main channel13and the metering channels15. When the diluted mixture is introduced into the metering channel15, a gas contained in the metering channel15is transferred into the reaction well5through the injection channel17. Due to the transfer of the gas into the reaction well5, a gas contained in the reaction well5is partially transferred into the reaction well air vent channels19and21. Furthermore, a gas contained in the channels between the reaction well air vent channel19and the bellows53is sequentially moved toward the bellows53(see open arrows inFIG. 10). Further, due to the injection of the diluted mixture into the liquid drain space29, a gas contained in the channels between the liquid drain space29and the bellows53is sequentially moved toward the bellows53(see open arrows inFIG. 10). As a result, the bellows53expands.

As shown inFIG. 11, the switching valve63is rotated to connect the syringe channel51cto the channel39afor air suction and to connect the air vent channel53bto the air vent channel39bfor the well for air suction. Then, the syringe51is driven in a suction direction to suck a gas contained in the well39for air suction into the channel in the switching valve63, the syringe channel51c, and the syringe51. At this time, the bellows53contracts due to the decompression of the well39for air suction (see open arrows inFIG. 11), since the well39for air suction is connected to the bellows53through the air vent channels39e,39d, and39b, the switching valve63, and the air vent channel53b.

As shown inFIG. 12, the switching valve63is rotated to connect the syringe channel51cto the channel13aand to connect the air vent channel53bto the channels23aand25aas in the case of a connection state shown inFIG. 10. Then, the syringe51is driven in an extrusion direction to send a gas contained in the channel in the switching valve63, the syringe channel51c, and the syringe51into the main channel13to purge the diluted mixture from the main channel13(see open arrows inFIG. 12). At this time, the diluted mixture remains in the metering channels15(see dots inFIG. 12) because the injection channels17do not allow the passage of the diluted mixture at a purge pressure applied to purge the diluted mixture from the main channel13. The purged diluted mixture is injected into the liquid drain space29. Further, due to the injection of the diluted mixture into the liquid drain space29, a gas contained in the channels between the liquid drain space29and the bellows53is sequentially moved toward the bellows53(see open arrows inFIG. 12). As a result, the bellows53expands.

As shown inFIG. 13, the switching valve63is rotated to connect the syringe channel51cto the channel39afor air suction and to connect the air vent channel53bto the air vent channel39bfor the well for air suction as in the case of a connection state shown inFIG. 11. Then, the syringe51is driven in a suction direction to suck a gas contained in the well39for air suction into the channel in the switching valve63, the syringe channel51c, and the syringe51. At this time, as in the case described with reference toFIG. 11, the bellows53contracts (see open arrows inFIG. 13).

As shown inFIG. 14, the switching valve63is rotated to connect the syringe channel51cto the channel13aand to connect the air vent channel53bto the channel25a. It is noted that the connection state shown inFIG. 14is different from those shown inFIGS. 10 and 12in that the liquid drain space29, to which the downstream end of the main channel13is connected, is not connected to the channel in the switching valve63. Then, the syringe51is driven in an extrusion direction. Since the downstream end of the main channel13is not connected to the bellows53, a pressure larger than the liquid introduction pressure and the purge pressure is applied to the inside of the main channel13. As a result, the diluted mixture in the metering channels15is injected into the reaction wells5through the injection channels17. After the completion of the injection of the diluted mixture into the reaction wells5, a gas contained in the main channel13is partially flown into the reaction wells5through the metering channels15and the injection channels17. At this time, a gas contained in the channels between the reaction wells5and the bellows53is sequentially moved toward the bellows53(see open arrows inFIG. 14), since the reaction wells5are connected to the bellows53through the reaction well air vent channels19and21, the air drain space31, the drain space air vent channel25a, and the air vent channel53b. As a result, the bellows53expands.

The switching valve63is returned to its initial state shown inFIG. 1Ato hermetically seal the wells, channels, and drain spaces provided in the reactor plate1. Then, the reaction wells5are heated by the temperature control system67to melt the wax9. As a result, the diluted mixture injected into each of the reaction wells5sinks below the wax9and, therefore, the diluted mixture is mixed with the reagent7so that a reaction occurs. As described above, by using the reactor plate1, it is possible to perform reaction processing in a closed system.

Alternatively, the wax9may be melted before the injection of the diluted mixture into the reaction wells5by heating the reaction wells5by the temperature control system67so that the diluted mixture is injected into the reaction wells5containing the melted wax9. In this case, the diluted mixture injected into each of the reaction wells5immediately sinks below the wax9, and is then mixed with the reagent7so that a reaction occurs. Even when the switching valve63is in the connection state shown inFIG. 14, the hermeticity of the reactor plate1is maintained by the bellows53. By returning the switching valve63to its initial state shown inFIG. 1Aafter the injection of the diluted mixture into the reaction wells5, it is possible to hermetically seal the wells, channels, and the drain spaces provided in the reactor plate1. It is noted that the switching valve63may be returned to its initial state shown inFIG. 1Aat any timing during the period from just after the injection of the diluted mixture into the reaction wells5until the end of the reaction between the diluted mixture and the reagent7, or may be returned to its initial state shown inFIG. 1Aafter the completion of the reaction between the diluted mixture and the reagent7.

As described above, by using the reactor plate1, it is possible to perform reaction processing in a closed system. In addition, it is also possible to maintain the hermeticity of the reactor plate1before and after reaction processing.

According to the present embodiment, grooves for forming the channels13,15,17,19,21, and23are provided in the channel base11, but the present invention is not limited to this embodiment. For example, grooves for forming all or part of these channels may be provided in the surface of the well base3.

FIG. 15is an expanded sectional view schematically showing a reaction well of a reactor plate according to another embodiment of the present invention and its vicinity. The reactor plate according to another embodiment of the present invention has the same structure as the reactor plate described above with reference toFIGS. 1A to 14except that a channel spacer is provided between the well base and the channel base.

On the well base3, a channel spacer73is provided to cover a region where the reaction wells5are arranged. On the channel spacer73, the channel base11and the channel cover33are further provided in this order. The channel spacer73is made of, for example, PDMS or silicone rubber. The thickness of the channel spacer73is, for example, from 0.5 to 5.0 mm. The channel spacer73has a projecting portion75projecting into each of the reaction wells5. The projecting portion75is substantially trapezoidal in cross section. For example, the proximal end of the projecting portion75has a width of 1.0 to 2.8 mm, and the distal end of the projecting portion75has a width of 0.2 to 0.5 mm. That is, the distal end of the projecting portion75is narrower than the proximal end of the projecting portion75. Further, the projecting portion75has a super-water-repellent surface. In this regard, it is noted that it is not always necessary to subject the surface of the projecting portion75to water-repellent treatment.

Further, in the channel spacer73, an injection channel77is provided at a position corresponding to each of the projecting portions75. The injection channel77is constituted from a through hole extending from the distal end of the projecting portion75to the surface of the channel spacer73where the projecting portion75is not provided. The injection channel77has an inner diameter of, for example, 500 μm. The opening of the injection channel77provided on the channel base11side is connected to the injection channel17provided in the channel base11. It is noted that the reactor plate according to another embodiment of the present invention is different from the reactor plate described above with reference toFIGS. 1A to 14in that the channel base11does not have a recess27.

Further the channel spacer73has a reaction well air vent channel79constituted from a through hole. The reaction well air vent channel79is provided to allow the reaction well5to communicate with the reaction well air vent channel19provided in the channel base11.

Although not shown inFIG. 15, the channel spacer73has through holes at positions corresponding to both ends of the main channel13, one end of each of the reaction well air vent channels21located on the air drain space31side, and both ends of each of the drain space air vent channels23and25to connect these channels13,21,23, and25to the wells29and31provided in the well base3and the channels23aand25a.

According to the embodiment of the present invention shown inFIG. 15, the end of the injection channel77on the opposite side from the injection channel17(i.e., the other end of the injection channel) is located at the tip of the projecting portion75which projects from the top inner surface of the reaction well5and, therefore, a liquid is easily dropped into the reaction well5through the injection channels17and77when injected into the reaction well5.

Further, by placing the tip of the projecting portion75in the vicinity of the side wall of the reaction well5so that when a liquid passes through the injection channel77and is then discharged from the tip of the projecting portion75, a droplet of the liquid formed at the tip of the projecting portion75can come into contact with the side wall of the reaction well5, it is possible to inject the liquid into the reaction well5along the side wall of the reaction well5, thereby making it possible to more reliably inject the liquid into the reaction well5. However, the projecting portion75may be provided at a position which does not allow a droplet formed at the tip of the projecting portion75to be brought into contact with the side wall of the reaction well5.

FIG. 16is an expanded sectional view schematically showing a reaction well of a reactor plate according to another embodiment of the present invention and its vicinity.

The reactor plate according to another embodiment of the present invention shown inFIG. 16is different from the reactor plate described above with reference toFIG. 15in that a projecting portion81is further provided in the reaction well5. The tip of the projecting portion81is located under the tip of the projecting portion75. By providing the projecting portion81, it becomes easy to guide a droplet formed at the tip of the projecting portion75into the reaction well5. The projecting portion81becomes particularly effective by subjecting the surface of at least the tip of the projecting portion81to hydrophilic treatment.

FIG. 17is an expanded sectional view schematically showing a reaction well of a reactor plate according to another embodiment of the present invention and its vicinity.

The reactor plate according to another embodiment of the present invention shown inFIG. 17is different from the reactor plate described above with reference toFIG. 16in that a stepped portion83and a linear projecting portion85, which is provided on the top surface of the stepped portion83in such a manner that a space is left between the tip of the linear projecting portion85and the top surface of the reaction well5, is further provided in the side wall of the reaction well5. The stepped portion83and the linear projecting portion85are circular when viewed from above. The tip of the linear projecting portion85is provided in such a manner that a space is left between the tip of the linear projecting portion85and the side wall of the reaction well5.

By providing the linear projecting portion85in such a manner that a space is left between the tip of the linear projecting portion85and the top surface of the reaction well5and between the tip of the linear projecting portion85and the side wall of the reaction well5, it is possible to prevent a liquid contained in the reaction well5from reaching the top surface of the reaction well5through the side wall of the reaction well5. The linear projecting portion85becomes particularly effective by subjecting the surface of at least the tip of the linear projecting portion85to water-repellent treatment.

The stepped portion83and the linear projecting portion85shown inFIG. 17can also be applied to the embodiment shown inFIG. 15.

In each of these various embodiments described above with reference toFIGS. 15 to 18, grooves for forming the channels13,15,17,19,21, and23are provided in the channel base11, but the present invention is not limited to these embodiments. For example, grooves for forming all or part of these channels may be provided in any one of the surfaces of the channel spacer73located on the channel base11side, the surface of the channel spacer73located on the well base3side, and the surface of the well base3.

A part of the cylinder51aof the syringe51may be constituted from a part of the switching valve63.

FIGS. 18A,18B, and18C show a reactor plate according to another embodiment of the present invention. More specifically,FIG. 18Ais a schematic plan view of the reactor plate according to another embodiment of the present invention,FIG. 18Bis a schematic sectional view taken along the A-A line inFIG. 18A, which further includes the sectional views of the metering channel15, the injection channel17, the reaction well air vent channels19and21, the liquid drain space29, the air drain space31, and the bellows53, andFIG. 18Cis an expanded sectional view schematically showing a syringe51, the bellows53and their vicinity.FIGS. 19A,19B, and19C are schematic exploded views of a switching valve95. More specifically,FIG. 19Ashows a plan view and sectional views of a sealing plate89,FIG. 19Bshows a plan view and sectional views of a rotor upper91, andFIG. 19Cshows a plan view and sectional views of a rotor base.

In the reactor plate in the embodiment shown inFIG. 18A, the syringe87has a cylinder87amade of, for example, a resin material such as polypropylene or polycarbonate, and the cylinder87ais integrally molded with the rotor upper91of the switching valve95.

The syringe87is constituted from the cylinder87aprovided in a through hole formed in the well base3and the well bottom55, a plunger87bplaced in the cylinder87a, and a cover body87d.

The cover body87dhas flexibility in the sliding direction of the plunger87b, and is connected to the cylinder87aand the plunger87b. The cover body87dis provided to create a sealed space87eto hermetically cut off a part of an inner wall of the cylinder87ato be brought into contact with the plunger87bfrom an atmosphere outside the cylinder87a. The sealed space87eis enclosed with the cylinder87a, the plunger87b, and the cover body87d.

One end of the cover body87dconnected to the cylinder87ais hermetically fixed to the upper end of the cylinder87aby a cylinder cap87f. On the other hand, the other end of the cover body87dconnected to the plunger87bis hermetically connected to the upper surface of the plunger87bby an adhesive. However, a method for connecting the cover body87dto the cylinder87aand the plunger87bis not limited to the method described above, and the connecting positions of the cover body87dare not limited to those described above. The plunger and the cover body may be integrally molded. The plunger and the cover body can be integrally molded using, for example, silicone rubber.

As described above, the cover body87dis connected to the cylinder87aand the plunger87bto create a sealed space87eenclosed with the cylinder87a, the plunger87b, and the cover body87dand, therefore, it is possible to prevent the entry of foreign matter from outside through a gap between the cylinder87aand the plunger87b. In addition, it is also possible to prevent the leakage of a liquid through the gap between the cylinder87aand the plunger87b, thereby preventing the pollution of an outside environment with the liquid. It is to be noted that as described above, since the cover body87dhas flexibility in the sliding direction of the plunger87b, the plunger87bcan be slidably moved.

Referring toFIGS. 19A,19B, and19C, the syringe air vent channel53cand the switching valve95will be described.

The switching valve95is constituted from the disk-shaped sealing plate89, rotor upper91, and rotor base93, and is attached to the well bottom55by means of a lock65.

The sealing plate89has a through hole89a, a through groove89b, and a through hole89c. The through hole89ais provided in the vicinity of the peripheral portion of the sealing plate89, and is connected to any one of the channels13a,35a,37a, and39a. The through groove89bis provided inside the through hole89aand on a circle concentric with the sealing plate89, and is connected to at least two of the channels23a,25a,35b,37b,39b, and53b. The through hole89cis provided at the center of the sealing plate89to insert the cylinder87athereinto. On the surface of the sealing plate89opposed to the well bottom55, a fluorine resin layer (not shown) is provided.

The rotor upper91has the cylindrical cylinder87a, a through hole91a, a groove91b, a through hole91c, and a through hole91d. The cylinder87ais provided on one surface of the rotor upper91so as to be located in the central portion of the rotor upper91. The through hole91ais provided at a position corresponding to the position of the through hole89aprovided in the sealing plate89. The groove91bis provided in the surface of the rotor upper91so as to correspond to the through groove89bprovided in the sealing plate89. The through hole91cis provided in the groove91b, and the through hole91dis provided at the center of the rotor upper91. The through hole91dis located at the bottom of the cylinder87aand constitutes a discharge port of the cylinder87a.

In the rotor upper91, the syringe air vent channel53cconstituted from a through hole extending from the upper end surface of the cylinder87ato the back surface of the rotor upper91is also provided. In the upper end surface of the cylinder87a, a notch extending from the surface of the inner wall of the cylinder87ato the syringe air vent channel53cis provided. As shown inFIG. 18C, the notch allows the sealed space87eto communicate with the syringe air vent channel53cin a state where the upper end surface of the cylinder87ais covered with the cover body87d.

The rotor base93has a groove93aand a groove93bin its surface to be bonded to the back surface of the rotor upper91. The groove93ais provided to connect the through hole91aand the through hole91dprovided in the rotor upper91to each other, and the groove93bis provided to connect the syringe air vent channel53cand the through hole91cprovided in the rotor upper91to each other.

As shown inFIG. 18B, the sealing plate89, the rotor upper91, and the rotor base93constituting the switching valve95are superposed so that the cylinder87ais inserted into the through hole89cof the sealing plate89.

The through hole91dof the rotor upper91constituting a discharge port of the cylinder87ais connected to the through hole89aof the sealing plate89through the groove93aof the rotor base93and the through hole91aof the rotor upper91.

The sealed space87e(seeFIG. 18C) is connected to the through groove89bof the sealing plate89through the syringe air vent channel53c, the groove93bof the rotor base93, the through hole91cand the through groove91bof the rotor upper91.

Referring toFIGS. 18A,18B,18C,19A,19B, and19C, channel connection will be described.

By rotating the switching valve95, the through hole91dof the rotor upper91constituting a discharge port of the cylinder87ais connected to any one of the channels13a,35a,37a, and39athrough the groove93a, the through hole91a, and the through hole89a.

Further, at the same time as the through hole91dis connected to any one of the channels13a,35a,37a, and39a, the air vent channel53bis connected to at least any one of the channels23a,25a,35b,37b, and39bthrough the through grooves89band91b. At this time, the sealed space87eis connected to the air vent channel53bthrough the cylinder air vent channel53c, the groove93b, the through hole91c, and the through grooves89band91b.

According to this embodiment, it is possible to eliminate the necessity to provide a channel between the syringe87and the switching valve95, thereby simplifying the channel configuration of the reactor plate.

In general, in a case where a channel has a joint, there is a possibility that a liquid or a gas will leak through the joint or a liquid will be accumulated in the joint. However, according to this embodiment, since the cylinder87aand the rotor upper91are integrally molded, there is no joint between the syringe87and the switching valve95and, therefore, liquid leakage, gas leakage, and liquid accumulation do not occur between the syringe87and the switching valve95.

In general, in a case where liquid accumulation occurs in the joint of a channel, there is a fear of reduction in the volume of a liquid to be fed through the channel, carry-over of a liquid accumulated in the joint of the channel into another liquid fed through the channel or a contamination of another liquid fed through the channel with a liquid accumulated in the joint of the channel, or fluctuation in the concentration of a liquid fed through the channel. However, according to this embodiment, since there is no joint between the syringe87and the switching valve95, such a fear can be eliminated.

As shown inFIGS. 1A,1B, and1C, in a case where the channel51cis provided between the syringe51and the switching valve63to arrange the syringe above the switching valve, the cylinder51acannot be formed in a portion where the channel51cis provided. However, since the reactor plate according to the embodiment shown inFIGS. 18A,18B, and18C does not need to have a channel between the syringe87and the switching valve95, the capacity of the cylinder87acan be made larger than that of the cylinder51aeven when the cylinder51aand the cylinder81ahave the same two-dimensional size.

In a case where the cylinder87ais formed to have the same capacity and two-dimensional size as the cylinder51a, the level of the upper end surface of the cylinder87acan be made lower than that of the cylinder51a. On the other hand, in a case where the cylinder87ais formed to have the same capacity and the upper end surface level as the cylinder51a, the two-dimensional size of the cylinder87acan be made smaller than that of the cylinder51a.

For example, even in a case where the upper end surface of the cylinder51aneeds to be located at a higher level than the upper surface of the entire reactor plate1due to limitations on the two-dimensional size of the entire reactor plate1, since the level of the upper end surface of the cylinder87acan be made lower than that of the cylinder51awhile the capacity and two-dimensional size of the cylinder87aremain the same as the cylinder51a, it is possible to allow the upper end surface of the cylinder87ato be located at the same or lower level than the upper surface of the entire reactor plate1. This makes it possible to eliminate problems caused by the cylinder whose upper end surface is located at a higher level than the upper surface of the entire reactor plate, such as difficulty in stacking two or more reactor plates on top of each other for storage and increase in the size of a package of the reactor plate.

By making the two-dimensional size of the cylinder87asmaller than that of the cylinder51awhile the capacity of the cylinder87aremains the same as the cylinder51a, it is also possible to reduce the two-dimensional size of the entire reactor plate1.

Although the present invention has been described above with reference to the various embodiments, the present invention is not limited to these embodiments. The shape, material, position, number, and size of each component and the channel configuration of the reactor plate in the above description are merely examples, and various changes can be made without departing from the scope of the present invention defined in claims.

For example, the bellows53connected to the air vent channel53bmay have another structure as long as it is a variable capacity member whose internal capacity is passively variable. Examples of such a bellows53having another structure include a bag-shaped one made of a flexible material and a syringe-shaped one.

The reactor plate according to the present invention does not always need to have a variable capacity member such as a bellows53. Further, in a case where a liquid such as a reagent is not previously contained in the well35,37, or39, the air vent channel thereof does not always need to partially have the channel35e,37e, or39econstituted from a narrow hole.

In each of the above embodiments, the air vent channels35b,37b, and39b, which communicate with the wells35,37, and39provided as sealed wells, are connected to the air vent channel53bthrough the switching valve63, but may be directly connected to the outside of the reactor plate or a variable capacity part such as a bellows53. Each of the wells35,37, and39may be sealed by using an openable and closable cap.

In each of the above embodiments, the well base3is constituted from one component, but may be constituted from two or more components.

The reagent contained in the reaction well5may be a dry reagent.

It is noted that the sample well35and the reaction well5do not always need to previously contain a reagent.

In each of the above embodiments, the reagent well37contains dilution water49, but may contain a reagent instead of the dilution water49.

The well base3may further have a gene amplification well for carrying out gene amplification reaction. For example, the empty reagent well37may be used as a gene amplification well.

By previously placing a reagent for gene amplification reaction in the reaction well5, it is possible to carry out gene amplification reaction in the reaction well5.

In a case where a liquid to be introduced into the main channel13contains a gene, a probe which reacts with the gene may be previously placed in the reaction well5.

In each of the above embodiments, the syringe51is placed on the switching valve63. However, the position of the syringe51is not limited to a position on the switching valve63, and the syringe51may be placed at any position.

In each of the above embodiments, the rotary switching valve63is used as a switching valve. However, a switching valve for use in the reactor plate according to the present invention is not limited thereto, and various channel switching valves can be used. The reactor plate according to the present invention may have a plurality of switching valves.

In each of the above embodiments, a liquid filling the metering channel15is injected into the reaction well5through the injection channel17by applying a pressure to the inside of the main channel13after air purge, but the reaction processing method according to the present invention is not limited to such a method. For example, a liquid filling the metering channel15may be injected into the reaction well5through the injection channel17by creating a negative pressure in the reaction well air vent channel21and then in the reaction well5. In this case, it is necessary to change the channel configuration of the reactor plate so that a negative pressure can be created in the reaction well air vent channel21by using the syringe51. Alternatively, another syringe may be additionally prepared. In this case, a positive pressure is created in the main channel13and a negative pressure is created in the reaction well5to inject the liquid into the reaction well5.

In each of the above embodiments, one main channel13is provided, and all the metering channels15are connected to the main channel13. However, the channel configuration of the reactor plate according to the present invention is not limited thereto. For example, a plurality of main channels may be provided. In this case, one or more metering channels may be connected to each of the main channels.

In the reactor plate according to the present invention, the main channel can be hermetically sealed. In this regard, the main channel may be hermetically sealed by, for example, allowing both ends of the main channel to be openable and closable. The phrase “allowing both ends of the main channel to be openable and closable” includes a case where each end of the main channel is connected to another space, and the end of the space located on the opposite side from the main channel is openable and closable. In the case of each of the above embodiments, such ‘another space’ corresponds to, for example, the channel13a, the liquid drain space29, the drain space air vent channel23, or the channel23a.

In the reactor plate according to the present invention, the reaction well air vent channel can be hermetically sealed. In this regard, the reaction well air vent channel may be hermetically sealed by, for example, allowing the end of the reaction well air vent channel located on the opposite side from the reaction well to be openable and closable. The phrase “allowing the end of the reaction well air vent channel located on the opposite side from the reaction well to be openable and closable” includes a case where the end of the reaction well air vent channel located on the opposite side from the reaction well is connected to another space, and the end of the space located on the opposite side from the reaction well air vent channel is openable and closable. In the case of each of the above embodiments, such ‘another space’ corresponds to, for example, the air drain space31, the drain space air vent channel25, or the channel25a.

In the case of such an aspect, a liquid is introduced into the main channel and the metering channels, and next, the liquid is purged from the main channel, and further, the liquid remaining in the metering channels is injected into the reaction wells, and thereafter both ends of the main channel and one end of the reaction well air vent channel located on the opposite side from the reaction well are closed to hermetically seal the main channel and the reaction well air vent channel.

The present invention can be applied to measurements of various chemical and biochemical reactions.