JP2003279584A - Chemical analysis apparatus and method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a chemical analysis apparatus that do not evaporate samples that are dispensed into a reaction bath, can seal the reaction bath without allowing gas to remain, and has improved determination properties and safety, and a dispensing method. <P>SOLUTION: The chemical analysis apparatus comprises a container-like member 8 where the reaction bath 5a for dispensing the samples is arranged, a cover member 11 for covering a surface at a side for dispensing the samples of the container-like member 8, a heating cooler 7 for maintaining temperature inside the reaction bath 5a at the freezing point or less the samples, and a block 12a for placement for placing the container-shaped member 8 via the heating cooler 7. In this case, the volume of the samples to be dispensed is expressed by V<SB>1</SB>≤V≤aV<SB>1</SB>when the volume of the samples in freezing is set to V, the volume of the reaction bath 5a is set to V<SB>1</SB>, and the coefficient of expansion when a sample liquid is frozen to a solid is set to a (=solid volume/ liquid volume). <P>COPYRIGHT: (C)2004,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a chemical analyzer and a dispensing method which are excellent in quantitativeness and safety.

[0002]

2. Description of the Related Art In recent years, μ-TAS (Micro Total Analys
There is proposed a chemical / biochemical analysis integrated system called is System) in which functional parts such as liquid feeding, mixing, reaction, and analysis are integrated on a chip such as glass or silicon of several cm square. FIG. 13 shows one of conventional examples of the μ-TAS. As shown in FIG. 13, in the μ-TAS, the sample channel 51 is formed in the glass chip 80, and the sample (reagent) is mixed, reacted, and detected in the sample channel 51.
Conduct preliminary experiments such as drug discovery and medical diagnosis. When the sample is injected from the sample injection section 62, the sample passes through the sample flow path 51 and is mixed in the mixing section 61a. Then, the chemical reaction of the sample occurs in the reaction section 61b, and the separated sample is separated in the separation section 61c. The sample after the reaction is detected by the detection unit 61d, and the unnecessary sample is collected by the waste liquid unit 60. Since the μ-TAS can significantly reduce the amount of samples and samples compared to the conventional integrated analysis system, it is expected to reduce the throughput time and waste liquid.

Further, there has been proposed a microreactor in which these device functions are built in a minute space such as a microchip and a plurality of more complicated chemical reactions, drug discovery and the like can be performed at one time. In these microreactors, handling of minute amounts of chemicals is important. For example, when dispensing a chemical solution or the like into a micro reaction tank on a chip or spotting, a means such as a syringe, an inkjet mechanism, or a spotter is used.

[0004]

It is difficult to handle a chemical solution with a μ-TAS chip because the solution volume is very small, and it is difficult to secure quantitativeness because the solution volume decreases due to evaporation or adhesion to a wall. The conventional method has hindered the control of the concentration of the drug solution.

For example, when the same chemical solution is dispensed into a plurality of reaction tanks (eg, the sample injection section 62 in FIG. 13), the chemical solution evaporates, so that the amount of the chemical solution in the reaction tank first injected. There is a possibility that the amount of chemical liquid in the reaction tank injected last may be different. This may cause a problem in quantification. Further, when dispensing a plurality of chemicals, evaporation may change the mixing ratio of the plurality of chemicals.

Further, in the micro PCR device, after the chemical liquid is dispensed into the container, the cover is covered and the inside is heated and cooled. However, the chemical liquid may evaporate after dispensing and the liquid volume may become smaller than the reaction tank capacity. . At this time, if the cover is left as it is, an air layer remains inside the reaction tank, so that there is a risk that the air expands and bursts when heated.

Therefore, in the present invention, a chemical analyzer which is excellent in quantitativeness and safety, in which the sample dispensed in the reaction tank is not evaporated and the reaction tank can be sealed without leaving a gas, and The purpose is to provide a dispensing method.

[0008]

In order to achieve the above object, a first feature of the present invention is (a) a container-shaped member in which a reaction tank for injecting a sample is arranged, and (b) the container-shaped member. A cover member that covers the surface of the member on which the sample is injected, (c) a heating / cooling device that keeps the internal temperature of the reaction chamber below the freezing point of the sample, and (d) a container-shaped member is placed via the heating / cooling device. The chemical analysis device is provided with a mounting block for performing the chemical analysis device. Here, the “chemical analysis device” refers to a microreactor in which various chemical analysis-related functions are integrated. This chemical analyzer also has functions of μ-TAS and nucleic acid amplifier.

According to the chemical analysis device of the first feature,
Since the reaction vessel is sealed with the cover member to the sample in the solidified state, the sample dispensed in the reaction vessel does not evaporate, and
The reaction tank can be sealed without leaving gas.

A second feature of the present invention is (a) a mounting block in which a reaction tank for injecting a sample is arranged, and (b)
A sealing lid having a gas inlet / outlet hole for sealing the surface of the reaction tank for injecting the sample, (c) a sensor for detecting the state of the sample provided in the sealing lid, and (d) penetrating the sealing lid, A gist of the chemical analysis device is an injection needle for injecting a sample into a reaction tank. The injection needle of the chemical analysis device according to the second feature is (e) a plurality of connectors for injecting a sample,
(V) A plurality of valves may be provided for connecting / disconnecting the flow of the sample between the connector and the reaction tank.

According to the chemical analysis device of the second feature,
The sample dispensed into the reaction tank does not evaporate and the reaction tank can be sealed without leaving any gas. Further, the sample after analysis can be easily extracted.

A third feature of the present invention is: (a) a reaction tank for injecting a sample, which is composed of a container-shaped member having a first adhesive member and a second adhesive member sandwiched between the members. ,
(B) a mounting block having wells for arranging a reaction tank, (c) a third bonding member that contacts the second bonding member, and a fourth bonding member that faces the third bonding member. It is a gist of the present invention to provide a chemical analyzer including a bonding member and a sealing lid that seals a surface of a reaction tank, which is composed of a cover member sandwiched around the surface, into which a sample is injected. The second bonding member and the third bonding member of the chemical analysis device according to the third feature may be bonded with an adhesive. The second bonding member and the third bonding member of the chemical analysis device according to the third feature may be provided with a concave portion and a convex portion, respectively, and may be joined by fitting the concave portion and the convex portion. .

According to the chemical analysis device of the third feature,
The sample dispensed into the reaction tank does not evaporate and the reaction tank can be sealed without leaving any gas. Further, even a reaction tank and a closed lid made of a material that is difficult to adhere can be easily adhered by an adhesive member.

The fourth feature of the present invention is (a) placing the container-shaped member on the mounting block, and (b) controlling the temperature inside the reaction vessel of the container-shaped member below the freezing point of the sample. And (c) a step of dispensing a sample into a reaction tank, (d) a step of adhering a cover member to the surface of the container-shaped member on which the sample is injected, and (e) a step of adhering The gist is a dispensing method including the step of controlling the temperature inside the tank to be equal to or higher than the melting point of the sample. In the dispensing step of the dispensing method according to the fourth feature, the dispensing rate of the sample may be the same as or slower than the rate at which the sample solidifies. Also, in the dispensing step of the dispensing method according to the fourth feature, the volume of the sample when solidified is V, the volume of the reaction tank is V1, and the expansion coefficient when the liquid of the sample is solidified is a ( = Solid volume / liquid volume), V ₁
≦ V ≦ a · V ₁ may be satisfied.

According to the dispensing method of the fourth feature, since the reaction vessel is sealed by the cover member to the sample in the solidified state, the sample dispensed in the reaction vessel does not evaporate and the gas is vaporized. It is possible to seal the reaction tank without leaving.

The fifth feature of the present invention is (a) a step of opening a valve and a gas inlet / outlet hole of an injection needle for injecting a sample into a reaction tank, and (b) a reaction of the sample from a connector of the injection needle. And (c) when it is detected that the sample has reached the sealing lid for sealing the sample injection surface of the reaction tank, the step of closing the valve and the gas inlet / outlet hole is included. The gist is that it is an injection method.

According to the dispensing method according to the fifth feature, the sample dispensed in the reaction tank does not evaporate, and the reaction tank can be sealed without leaving any gas. Further, the sample after analysis can be easily extracted.

A sixth feature of the present invention is (a) a reaction vessel for injecting a sample, which is composed of a container-shaped member having a first adhesive member and a second adhesive member sandwiched in between. A step of dispensing a sample; (b) a reaction tank, a third bonding member that contacts the second bonding member, and a fourth bonding member that faces the third bonding member, The gist of the dispensing method is to include a step of fitting a closed lid made of a sandwiched cover member, and (c) a step of disposing a reaction tank on a mounting block. The fitting step of the dispensing method according to the sixth aspect may include a step of joining the second adhesive member and the third adhesive member with an adhesive.
Further, the fitting step of the dispensing method according to the sixth feature may include a step of fitting the second bonding member and the third bonding member by fitting the concave portion and the convex portion respectively. .

According to the dispensing method of the sixth feature, the sample dispensed in the reaction tank is not evaporated and the reaction tank can be sealed without leaving any gas. Further, even a reaction tank and a closed lid made of a material that is difficult to adhere can be easily adhered by an adhesive member.

[0020]

BEST MODE FOR CARRYING OUT THE INVENTION Next, first to third embodiments of the present invention will be described with reference to the drawings. In the following description of the drawings, the same or similar parts are denoted by the same or similar reference numerals. However, it should be noted that the drawings are schematic.

(First Embodiment) As shown in FIG. 1A, the chemical analyzer according to the first embodiment is a container-shaped member 8 in which a reaction tank 5a for injecting a sample is arranged. When,
A cover member 11 that covers the surface of the container-shaped member 8 on the side where the sample is injected, a heating / cooling device 7 that keeps the temperature inside the reaction tank 5a below the freezing point of the sample, and the container-shaped member 8 through the heating / cooling device 7 And a mounting block 12a for mounting the same. Here, the volume of the sample to be dispensed is V, the volume of the sample when it is solidified, V _{1 is} the volume of the reaction tank 5a, and a coefficient of expansion when the liquid of the sample is solidified is a (= solid volume / liquid). Volume), V ₁ ≦ V ≦ a · V ₁ . When the volume V when solidified is equal to V ₁ , the volume of the solidified sample is equal to the volume V _{1 of the} reaction tank 5a. When the volume V when coagulated becomes equal to a · V ₁ , the volume of the melted sample becomes equal to the volume V _{1 of the} reaction tank 5a.

The container-shaped member 8 and the cover member 11 are
It is made of a material such as polypropylene having a thickness of about 50 μm, which has strong chemical resistance and is flexible, such as polypropylene. For example, glass epoxy resin, fluororesin such as polypropylene (PP) and polytetrafluoroethylene (PTFE),
It is made of a glass material such as quartz, a semiconductor material such as silicon, or a metal. Here, examples of the metal include nichrome, hastelloy, mnemonic, inconel, and stainless steel, which have low thermal conductivity. The mounting block 12a includes a heating / cooling device 7. As the heating / cooling device 7, a structure in which a Peltier element, a heat pipe, or the like is fed back using a temperature detecting element such as a thermocouple can be used.

FIG. 1B is a view of the chemical analysis device according to the first embodiment seen from above with the cover member 11 removed. The length L1 of the chemical analyzer is, for example, 10 to 3
The diameter d of the reaction tank 5a is preferably 1 μm to 5 mm. The diameter may be 5 mm or more, but the integration density is reduced and the function as a microreactor is reduced.
If the processing technology permits, a diameter d of 1 μm or less is possible, but it is not realistic from the viewpoint of operability and analysis sensitivity in performing mixing, reaction, and analysis. Therefore, considering the integration density as a microreactor, analytical sensitivity, and ease of manufacturing,
It may be about 0 μm to 1 mm. More preferably 0.
If it is about 1 to 1 mm, the integration density is kept high and the operability is good.

According to the chemical analyzer of the first embodiment, the reaction vessel 5a is sealed by the cover member 11 in the solidified sample, so that the sample dispensed in the reaction vessel 5a evaporates. In addition, the reaction tank 5a can be sealed without leaving any gas.

Next, a method of dispensing using the chemical analyzer according to the first embodiment will be described.

(A) First, as shown in FIG. 2A, the container-shaped member 8 is placed on the mounting block 12a having the heating / cooling device 7. The container member 8 has a concave reaction tank 5a. At this time, the heating / cooling device 7 is temperature-controlled at a temperature below the freezing point of the sample. When the liquid sample 10b is injected into the reaction tank 5a from the dispensing needle 6 of the dispensing device 9, the liquid sample 10b is solidified. Here, the dispensing device 9 has an injection needle-shaped dispensing needle 6 at its tip, and the inside diameter of the dispensing needle 6 is, for example, about 0.1 mm and the outside diameter is about 0.3 mm. If the diameter of the reaction tank 5a is about 1 μm, a microcapillary having a fine structure similar to the probe of the scanning tunneling microscope may be used. The dispensing device 9 may have a structure in which a plurality of dispensing needles 6 are provided in accordance with the reaction tank 5a of the container-shaped member 8 and the sample is injected into the plurality of reaction tanks 5a by one dispensing. .

(B) Injecting the liquid sample 10b into the reaction tank 5
Then, as shown in FIG. 2B, the sample solidified,
It becomes the solid sample 10a. The injection speed of the drug solution is
It is necessary to be less than the speed. Injection rate and solidification rate
If the degree is the same, the sample can be injected efficiently. Also, the sample
The atmosphere may dry up as condensation may form when it solidifies.
Desirable to be dry. Therefore, nitrogen with a low dew point
(N_Two) Gas, hydrogen (H _Two) Gas or helium
It is preferably performed in a rare gas such as (He) gas. This and
The volume of the injected sample when it is solidified is V, the reaction tank
The volume of 5a is V₁, When the sample liquid is solidified into a solid
Let the expansion coefficient of a be a (= solid volume / liquid volume), then V₁
≤V≤a ・ V₁Inject a sample of the volume Therefore,
The upper surface of the solid sample 10a is the same as or above the upper surface of the reaction tank 5a.
It will be above the height.

(C) Next, as shown in FIG. 2 (c), the cover member 11 covers the upper surface of the solid sample 10a.
Cover. The cover member 11 is a flexible material that can be flexibly shaped along the upper surface of the solid sample 10a. Therefore, by adhering the cover member 11 and the container-shaped member 8 to each other, it is possible to prevent air from entering the reaction tank 5a. In particular, if the cover member 8 is covered with N ₂ gas, H ₂ gas, rare gas or the like using a glove box or a remote control system, it is possible to prevent air from entering the reaction tank 5a. As a method for adhering the container-shaped member 8 and the cover member 11, besides using an adhesive, heat, ultrasonic vibration, or the like may be used.

(D) Next, as shown in FIG. 2 (d), the heating / cooling device 7 heats the sample to a temperature not lower than the melting point and not higher than the boiling point to melt the sample and return it to the liquid sample 10b. At this time, although the volume of the liquid sample 10b is reduced, the cover member 11 and the container-shaped member 8 have a pliability property, and their shapes can be changed according to the volume of the sample. No air is mixed in 5a.

(E) After that, the liquid sample 10b is analyzed. For example, when nucleic acid amplification is performed assuming that the liquid sample 10b is a nucleic acid (DNA), the liquid sample 10b is heated, DNA is dissociated at about 95 ° C., annealing is performed at about 55 ° C., and replication is performed at about 70 ° C. And amplify.

(V) After the analysis, the container-shaped member 8 containing the liquid sample 10b is removed from the mounting block 12a. After that, only the container-shaped member 8 and the cover member 11 that are in direct contact with the sample may be discarded, and the mounting block 12a, the heating / cooling device 7 and the like may be repeatedly used.

According to the chemical analysis apparatus of the first embodiment, the cover member 11 seals the reaction tank 5a to the solidified sample, so that the sample dispensed into the reaction tank 5a evaporates. In addition, the reaction tank 5a can be sealed without leaving any gas.

(Second Embodiment) As shown in FIG. 3, a chemical analyzer according to a second embodiment is a mounting block in which reaction tanks 5b and 5c for injecting a sample 10 are arranged. 12b and a gas inlet / outlet hole 25, and a reaction tank 5b,
Sealing lid 21a for sealing the surface of 5c into which the sample 10 is injected
A sensor 24 for detecting the state of the sample 10 provided on the sealing lid 21a and the sealing lid 21a.
An injection needle 20 for injecting the sample 10 is provided in b and 5c.

The mounting block 12b is made of, for example, glass epoxy resin, fluororesin such as polypropylene (PP) or polytetrafluoroethylene (PTFE), quartz, silicon, metal or the like. Here, examples of the metal include nichrome, hastelloy, mnemonic, inconel, and stainless steel, which have low thermal conductivity. Mounting block 12b
Is equipped with reaction tanks 5b and 5c. The sealing lid 21a is
The reaction tanks 5b and 5c are sealed and provided with an injection needle 20 and a gas inlet / outlet hole 25.

The injection needle 20, as shown in FIG.
A plurality of connectors 27a, 27c for injecting 0 and a plurality of valves 26a, 26b, 26c for connecting / disconnecting the flow of the sample 10 between the connectors 27a, 27c and the reaction tank are provided. Further, the sample 10 from the first connector 27a to the first valve 26a, which is connected / disconnected by the first valve 26a,
Is connected / disconnected by the first flow path 28a and the second valve 26b, and the flow path of the sample 10 following the reaction tank from the second valve 26b is connected / disconnected by the second flow path 28b and the third valve 26c. The flow path of the sample 10 from the second connector 27c to the third valve 26c is the third flow path 28c.

The first valve 26a, the second valve 26b, and the third valve 26c connect / disconnect the flow of the sample 10 inside the injection needle 20. As the structure of this valve, for example,
A single-beam structure passive valve may be provided inside the injection needle 20 to control the flow of the sample 10. Alternatively, the flow of the sample 10 may be controlled by changing the inner diameter of the injection needle 20 using a shape memory alloy or a piezoelectric element. Further, an electromagnetic valve that opens and closes with the force of an electromagnet (solenoid) may be used.

The gas inlet / outlet hole 25 is closed when necessary, so that the reaction tanks 5b and 5c have a closed structure. As the structure of the gas inlet / outlet hole 25, for example, a shutter which is electrically or magnetically driven at an arbitrary timing may be used. Alternatively, the gas inlet / outlet hole 25 may be formed of a polymer absorber, and the gas inlet / outlet hole 25 may be closed by the sample 10 soaking into the polymer absorber. Further, as shown in FIG. 5, a passive stopper 29 may be used. FIG. 5A is a cross-sectional view of the sealing lid 21 a in which the gas inlet / outlet hole 25 is formed in a trapezoidal shape, and the stopper 29 is arranged inside the gas inlet / outlet hole 25. When the sample 10 is filled in the reaction tank and reaches the height of the closed lid 21a, the stopper 2
9 is pushed up by the sample 10. The reaction vessel can be sealed by bringing the stopper 29 into close contact with the inner wall of the gas inlet / outlet hole 25. In this case, the material of the stopper 29 may be one having a specific gravity smaller than that of the sample 10. Also, FIG.
(B) is the figure which looked at the stopper 29 from the reaction tank side.
In order not to fall into the inside of the reaction tank, a cross-shaped stopper is provided at the lower part of the gas inlet / outlet hole 25 so as to continue with the sealing lid 21a. Needless to say, this stopper is not limited to a cross shape as long as the stopper 29 does not drop into the reaction tank.

The sensor 24 measures the conditions such as the height, temperature and PH value of the sample inside the reaction tanks 5b and 5c. In FIG. 3, the sensor 24 is installed on the upper part of the sealing lid 21a, but it may be installed on the gas inlet / outlet hole 25 inside the sealing lid 21a. In order to automatically detect whether or not the sample 10 inside the reaction tanks 5b and 5c has reached the height of the sealing lid 21a, an electrode is installed on the sealing lid 21a and the electric resistance value between the electrodes can be monitored. Good. At the moment when the sample 10 reaches the height of the sealing lid 21a, the electric resistance value decreases. Further, in order to measure the temperature of the sample 10 inside the reaction tanks 5b and 5c, a thermocouple, a resistance temperature detector or the like may be used. or,
The PH value may be measured using the glass electrode method.

According to the chemical analyzer of the second embodiment, the sample dispensed in the reaction tanks 5b and 5c is not evaporated, and the reaction tanks 5b and 5c are sealed without leaving any gas. You can Further, the sample 10 after analysis can be easily extracted.

Next, a method of dispensing using the chemical analyzer according to the second embodiment will be described with reference to FIG. In the following description, the chemical analyzer shown in FIG. 3 is described as a specific example, but it is assumed that all the injection needles 20 shown in FIG. 3 have the structure shown in detail in FIG. To do.

(A) First, in step S101,
Open the valve. At this time, like the reaction tank Ab on the left side of FIG. 3, the first valve 26a and the second valve 26b are
Open and close the third valve 26c, the first connector 2
A flow path for the sample 10 connecting from 7a to the reaction tank 5b is formed. Like the reaction tank 5c on the right side, when the first valve 26a is closed and the second valve 26b and the third valve 26c are opened, a flow path for the sample 10 is formed from the second connector 28c to the reaction tank 5c. Also, the gas inlet / outlet hole 25 is left open. As a result, the sample 10 becomes the reaction tank 5b,
When poured into 5c, the gas inside the reaction tanks 5b and 5c can escape to the outside of the reaction tanks 5b and 5c through the gas inlet / outlet hole 25.

(B) Next, in step S102,
Supply the sample to be dispensed to the connector. For example, in the reaction tank 5b on the left side of FIG.
Supply to. One kind of sample may be supplied in this manner, or the first valve 26a and the second valve 26b shown in FIG.
All the third valves 26c are opened, and the first connector 2
Samples may be supplied to both 7a and the second connector 27c, and a plurality of types of samples 10 may be supplied by connecting the channels of multiple inputs and one output.

In FIG. 6, it has been described that the valve and the gas inlet / outlet hole 25 are opened and then the sample is supplied to the connector. However, after the sample is supplied to the connector, the valve and the gas inlet / outlet hole 25 are opened. It doesn't matter.

(C) Next, in step S103,
The sample is supplied to the reaction tank. For example, in the reaction tank 5b on the left side of FIG. 3, the sample 10 is fed from the first connector 27a to the reaction tank 5b.
supply to b. At this time, the state of the sample 10 in the reaction tank 5b is constantly monitored by the sensor 24 installed on the closed lid 21a.

(D) In step S104, the sensor 24 detects whether the sample 10 has reached the height of the sealing lid 21a. If detected, the process proceeds to step S105, and the gas inlet / outlet hole 25 is closed. And FIG.
In (2), at least the second valve 26b is closed to shut off the flow path of the sample 10. If not detected, the process returns to step S103, and the sample is continuously supplied to the reaction tank.

(E) Next, in step S106,
The sample 10 is analyzed. For example, if the sample 10 is a nucleic acid (DN
In the case of performing nucleic acid amplification assuming A), the sample 10 is heated, the DNA is dissociated at about 95 ° C., the DNA is annealed at about 55 ° C., and the replication is performed at about 70 ° C. for amplification.

(V) After the analysis is completed, step S10
At 7, the valve is opened. If necessary, the gas inlet / outlet hole 25 is also opened. For example, in the reaction tank 5b on the left side of FIG. 3, the first valve 26a and the second valve 2
6b is opened.

(G) Next, in step S108,
The sample 10 is pulled out to the connector. For example, in the reaction tank 5b on the left side of FIG. 3, the sample 10 is extracted to the first connector 27a. The connector for extracting the sample 10 does not have to be the connector that supplied the sample. For example, the sample 10 in the reaction tank 5b on the left side of FIG. 3 is supplied from the first connector 27a,
It may be pulled out to the second connector 27c. In this case, the second valve 26b and the third valve 26c are kept open.

Here, in the dispensing method according to the second embodiment, the sample 10 inside the injection needle 20 is transferred to the reaction tanks 5b and 5b.
The force sent to the inside of c will be described. Reaction tanks 5b, 5
injection needle 20 that can be dispensed in c, for example, outer diameter 0.2 mm,
Considering the injection needle 20 having an inner diameter of 0.1 mm, the surface tension of the inner side surface of the injection needle 20 has a greater effect than the gravity applied to the sample 10. Therefore, the sample 10 is supplied to the connector only by supplying the sample 10 to the connector. Does not flow down inside. Therefore, it is necessary to add the output power to the sample 10 inside the injection needle 20. As this method, for example, a sample having a diameter slightly smaller than the inner diameter of the injection needle 20 may directly deliver the sample inside the injection needle 20. A pump (not shown), a tube (not shown), etc. may be connected to the upper portion of the connector, and positive pressure may be applied to the connector to generate the sample output. Further, when the gas inlet / outlet hole 25 is in an open state, a pump, a tube or the like is connected to the upper portion of the gas inlet / outlet hole 25 and a negative pressure is applied to the gas inlet / outlet hole 25 to generate a sample output. May be. Alternatively, the solution inside the injection needle 20 may be delivered by the peristaltic movement of the injection needle 20.

On the contrary, when the sample existing in the reaction tanks 5b and 5c is extracted to the first connector 27a, the first valve 26a and the second valve 26b are opened and the third valve is opened.
With the valve 26c closed, a pump, a tube, etc. are connected to the upper portion of the first connector 27a, and the first connector 2
It is sufficient to generate a sample output by applying a negative pressure to 7a. Alternatively, when the gas inlet / outlet hole 25 is in an open state, a pump, a tube, or the like is connected to the upper portion of the gas inlet / outlet hole 25 and a positive pressure is applied to the gas inlet / outlet hole 25 to generate a sample output. May be. Furthermore, the sample inside the injection needle 20 may be sent out by the peristaltic movement of the injection needle 20.

In the chemical analysis device according to the second embodiment, the first flow path 28a has a second flow path 28b and a third flow path 2b.
Although the structure is branched into 8c, the structure may be such that the third flow path 28c is omitted and the flow path is not branched, or the number of the flow paths is increased and the structure has a plurality of flow paths. Of course.

Although the chemical analyzer according to the second embodiment is described as having the injection needle 20 fixed to the closed lid 21a, the injection needle 20 is removable and the sample 10 is injected. It may be inserted into the sealing lid 21a before.
In this case, the injection needle 20 is removed after the analysis of the sample 10 is completed and the sample 10 is extracted. Injection needle 20
By making the needle removable, the injection needle 20 can be easily replaced and washed.

According to the dispensing method according to the second embodiment, the sample dispensed in the reaction tanks 5b and 5c does not evaporate,
Moreover, the reaction tanks 5b and 5c can be sealed without leaving gas. Further, the sample 10 after analysis can be easily extracted.

(Third Embodiment) As shown in FIGS. 7 and 8, a chemical analyzer according to a third embodiment includes a plurality of reaction tanks 5d for analyzing a sample and a plurality of reaction tanks. A mounting block 12c having a well 31 for arranging 5d,
Each of the plurality of reaction tanks 5d is provided with a plurality of sealing lids 21b for sealing the surface into which the sample 10 is injected. The mounting block 12c is made of, for example, glass epoxy resin, polypropylene (PP) or polytetrafluoroethylene (PTFE).
Fluorine resin, quartz, silicon, metal, etc. Here, examples of the metal include nichrome, hastelloy, mnemonic, inconel, and stainless steel, which have low thermal conductivity. The sealing lid 21b is a lid that seals the reaction tank 5d containing the sample so as to prevent air from entering.

FIG. 9 is a sectional view showing the details of the structures of the reaction tank 5d and the sealing lid 21b. The chemical analysis device according to the third embodiment includes the first bonding member 38.
And a second bonding member 37, which is composed of a container-shaped member 8 whose periphery is sandwiched, and a reaction tank 5d for injecting the sample 10.
And a mounting block 12c having a well for arranging the reaction tank 5d, and a third contacting the second bonding member 37.
A sealing for sealing the surface of the reaction tank 5d, which includes the cover member 11 sandwiched by the bonding member 36 of No. 4 and the fourth bonding member 35 facing the third bonding member 36, into which the sample 10 is injected. And a lid 21b. Third adhesive member 36
Has a shape in which no air can be accumulated by taking a corner inside the reaction tank 5d. The container-shaped member 8 and the cover member 11 are 10 to 50 μm, like the chemical analysis device according to the first embodiment.
It is made of a flexible material such as polypropylene with a thickness of about m, which has a strong chemical reaction and is flexible. The container-shaped member 8 and the cover member 11 are materials that are relatively difficult to bond. In order to bond these, it is necessary to perform a chemical or physical treatment such as corona treatment, acid treatment, sandblast treatment, and primer treatment to obtain a surface suitable for adhesion. Therefore, the adhesive members 35, 36, 3 sandwiching the periphery of the container-shaped member 8 and the cover member 11.
The container-shaped member 8 and the cover member 11 are bonded by bonding 7 and 38. Adhesive members 35, 36, 37, 3
The material of No. 8 is metal, glass, resin other than Teflon (registered trademark) / polypropylene, etc., which are relatively easy to bond. Alternatively, a thin film of metal or resin formed on the surface of any member may be used as the bonding members 35, 36, 37, 38. As a method of forming a thin film, sputtering,
A suitable method such as vapor deposition or dipping may be used depending on the base material.

In FIG. 9, the third adhesive member 36 and the second adhesive member 37 are joined by an adhesive 39, so that the inside of the reaction tank 5d is sealed by the sealing lid 21b. When the sealing lid 21b is fitted in the reaction tank 5d, the lower surface of the cover member 11 and the sample in the reaction tank 5d are in close contact with each other due to surface tension, and the air in the reaction tank 5d can be eliminated. When the amount of the sample 10 in the reaction tank 5d is small, the cover member 11 having the flexibility can be lowered to remove the air. When the amount of the sample 10 is large, the excess sample 10 is removed from the outside of the reaction tank 5d. Air can be eliminated to eliminate it. Further, instead of the adhesive 39, the bonding surfaces of the third bonding member 36 and the second bonding member 37 are heated by a heater so that the third bonding member 36 and the second bonding member 37 are separated from each other. The reaction tank 5d may be sealed with the sealing lid 21b by welding and bonding. Further, as shown in FIG.
When the 1b is fitted, the third adhesive member 36 may be provided with a convex portion and the second adhesive member 37 may be provided with a concave portion, and the reaction tank 5d may be hermetically sealed so that these are fitted together.

Further, as shown in FIG. 11, a plurality of reaction tanks 5
Each d may include a blocking member 40, and when the sealing lid 21b is fitted into the reaction tank 5d, the extra sample 10 removed outside the reaction tank 5d may be received. The blocking member 40 also functions as a partition of the sample 10 from the adjacent reaction tank 5d.

According to the chemical analyzer of the third embodiment, the sample dispensed in the reaction tank 5d is not evaporated and the reaction tank 5d can be sealed without leaving any gas.
Even if the reaction tank 5d and the sealing lid 21b are made of a material that is difficult to bond, the bonding members 35, 36, 37, 38
Can be easily bonded.

Next, a method of dispensing using the chemical analyzer according to the third embodiment will be described with reference to FIG. (A) First, in step S201, the sample 10 is dispensed into the reaction vessel 5d by the dispenser. This dispensing device may be the same as the dispensing device described in the dispensing method according to the first embodiment.

(B) Next, in step S202,
The closed lid 21b is fitted into the reaction tank 5d. Then, the third adhesive member 36 sandwiching the lower periphery of the cover member 11 and the second adhesive member 37 sandwiching the upper periphery of the container-shaped member 8 are coupled. As the joining method, the third adhesive member 36 and the second adhesive member 37 are joined by using an adhesive 39. Alternatively, the third adhesive member 36 may be provided with a convex portion and the second adhesive member 37 may be provided with a concave portion, and these may be joined so as to be fitted to each other. As a result, the reaction tank 5d and the sealing lid 21b are sealed.

(C) Next, in step S203,
The reaction tank 5d sealed with the sealing lid 21b is placed in the well 31 of the mounting block 12c.

(D) Next, in step S204,
The sample 10 is analyzed. For example, if the sample 10 is a nucleic acid (DN
In the case of performing nucleic acid amplification assuming A), the sample 10 is heated, the DNA is dissociated at about 95 ° C., the DNA is annealed at about 55 ° C., and the replication is performed at about 70 ° C. for amplification.

(E) After the analysis, the container-shaped member 8 containing the sample 10 is removed from the mounting block 12c. After that, only the container-shaped member 8 and the cover member 11 that are in direct contact with the sample 10 may be discarded and the mounting block 12c or the like may be repeatedly used.

In the above dispensing method, the sample 10 was dispensed into the reaction tank 5d and then installed on the mounting block 12c. However, the sample 10 was dispensed after the reaction tank 5d was installed on the mounting block 12c. It doesn't matter.

According to the dispensing method according to the third embodiment, the sample dispensed in the reaction tank 5d does not evaporate, and
The reaction tank 5d can be sealed without leaving a gas. or,
Reaction tank 5d and sealing lid 21 made of material that is difficult to bond
Even if it is b, it can be easily bonded by the bonding members 35, 36, 37 and 38.

(Other Embodiments) Although the present invention has been described by the above embodiments, it should not be understood that the description and drawings forming a part of this disclosure limit the present invention. From this disclosure, various alternative embodiments, examples and operational techniques will be apparent to those skilled in the art.

For example, in the first embodiment of the present invention, the mounting block 12a includes the heating / cooling device 7,
This device may be applied to the second and third embodiments. That is, in the second and third embodiments, the mounting blocks 12b and 12c may be provided with a plurality of heating and cooling devices, and the temperature may be controlled for each of the reaction tanks 5b, 5c, and 5d.

The dispensing method according to the first embodiment may be applied to the dispensing method according to the third embodiment of the present invention. That is, in the third embodiment, the mounting block 12c is provided with the heating / cooling device 7, the sample 10 is dispensed while being solidified, and the sample 10 is sealed with the sealing lid 21b.
0 may be melted. Further, the blocking member 40 described in the third embodiment may be used in the chemical analysis device according to the first and second embodiments. As described above, it goes without saying that the chemical analyzers and the dispensing methods according to the first to third embodiments may be used in combination.

As described above, it goes without saying that the present invention includes various embodiments not described here. Therefore, the technical scope of the present invention is defined only by the matters specifying the invention according to the scope of claims reasonable from the above description.

[0070]

EFFECTS OF THE INVENTION According to the present invention, a chemical analysis excellent in quantitativeness and safety, in which the sample dispensed in the reaction vessel is not evaporated and the reaction vessel can be sealed without leaving a gas. An apparatus and a dispensing method can be provided.

[Brief description of drawings]

FIG. 1A is a cross-sectional view of a chemical analysis device according to a first embodiment. (B) is the figure which looked at the chemical-analysis apparatus which concerns on 1st Embodiment from the top.

FIG. 2 is a method for dispensing a sample into the chemical analyzer according to the first embodiment.

FIG. 3 is a sectional view of a chemical analysis device according to a second embodiment.

FIG. 4 is a cross-sectional view of an injection needle of the chemical analysis device according to the second embodiment.

FIG. 5A is a cross-sectional view of a hermetically-sealing lid of a chemical analysis device according to a second embodiment. (B) is the figure which looked at the airtight lid provided with the stopper from the reaction tank side.

FIG. 6 is a flowchart of a dispensing method according to a second embodiment.

FIG. 7 is a perspective view of a chemical analysis device according to a third embodiment.

FIG. 8 is a sectional view of a chemical analysis device according to a third embodiment.

FIG. 9 is a detailed cross-sectional view of the chemical analysis device according to the third embodiment.

FIG. 10 is another detailed cross-sectional view of the chemical analysis device according to the third embodiment.

FIG. 11 is a perspective view in which a blocking member is attached to the chemical analysis device according to the third embodiment.

FIG. 12 is a flowchart of a dispensing method according to a third embodiment.

FIG. 13 is a perspective view of a conventional μ-TAS.

[Explanation of symbols]

5a, 5b, 5c, 5d reaction tank 6 dispensing needle 7 heating and cooling device 8 Container-shaped member 9 dispensing device 10 samples 10a solid sample 10b Liquid sample 11 Cover member 12a, 12b, 12c Mounting block 20 injection needle 21a, 21b Sealing lid 24 sensors 25 Gas in / out hole 26a First valve 26b Second valve 26c Third valve 27 connectors 27a First connector 27c Second connector 28a First flow path 28b Second flow path 28c Third flow path 29 Stopper 31 wells 35 Fourth Adhesive Member 36 Third adhesive member 37 Second adhesive member 38 First adhesive member 39 Adhesive 40 Blocking member

   ─────────────────────────────────────────────────── ─── Continued front page    (72) Inventor Hajime Sudo             1st Komukai Toshiba-cho, Sachi-ku, Kawasaki-shi, Kanagawa             Inside the Toshiba Research and Development Center (72) Inventor Yasushi Fukuda             1st Komukai Toshiba-cho, Sachi-ku, Kawasaki-shi, Kanagawa             Inside the Toshiba Research and Development Center (72) Inventor Masahiro Kuwata             1st Komukai Toshiba-cho, Sachi-ku, Kawasaki-shi, Kanagawa             Inside the Toshiba Research and Development Center F term (reference) 2G058 BB12 BB14 BB26 CC02 CC19                       EA08

Claims (13)

[Claims] 1. A container-shaped member in which a reaction tank for injecting a sample is arranged, a cover member for covering a surface of the container-shaped member on a side for injecting the sample, and a temperature inside the reaction tank for the sample. A chemical analysis device comprising: a heating / cooling device kept below a freezing point; and a mounting block for mounting the container-shaped member via the heating / cooling device. 2. A mounting block in which a reaction tank for injecting a sample is disposed, a gas inlet / outlet hole, and a sealing lid for sealing a surface of the reaction tank into which the sample is injected, and the sealing lid. A chemical analyzer comprising: a sensor for detecting the state of the sample provided; and an injection needle that penetrates the closed lid and injects the sample into the reaction tank. 3. The injection needle includes a plurality of connectors for injecting the sample, and a flow of the sample between the connectors and the reaction tank is conducted / conducted.
The chemical analyzer according to claim 2, further comprising a plurality of valves for shutting off. 4. A first bonding member and a second bonding member,
A reaction tank for injecting a sample, which is composed of a container-shaped member with its periphery sandwiched, a mounting block having a well for disposing the reaction tank, and a third block in contact with the second bonding member. An adhesive member and a fourth lid member facing the third adhesive member, and a sealing lid for sealing the surface of the reaction tank, which is composed of a cover member and is filled with the sample, for sealing the sample injection surface. A chemical analyzer characterized by the following. 5. The chemical analysis device according to claim 4, wherein the second adhesive member and the third adhesive member are bonded by an adhesive. 6. The second adhesive member and the third adhesive member each include a concave portion and a convex portion, and are joined by fitting the concave portion and the convex portion. The chemical analyzer according to claim 4. 7. A step of mounting a container-shaped member on a mounting block, a step of controlling a temperature inside the reaction tank of the container-shaped member to be equal to or lower than a freezing point of a sample, and a sample is dispensed into the reaction tank. And a step of adhering a cover member to the surface of the container-shaped member on which the sample is injected, and a step of controlling the temperature inside the reaction tank to be equal to or higher than the melting point of the sample after the adhering step. Dispensing method characterized by. 8. The dispensing method according to claim 7, wherein in the dispensing step, a dispensing rate of the sample is equal to or less than a rate at which the sample solidifies. 9. In the step of dispensing, the volume of the sample when solidified is V, the volume of the reaction tank is V ₁ ,
The coefficient of expansion when the sample is solidified from liquid to solid is a
If (= solid volume / liquid volume), V ₁ ≦ V ≦ a · V
_The dispensing method according to claim 7, wherein the dispensing method is 1. 10. A step of opening a valve and a gas inlet / outlet hole of an injection needle for injecting a sample into a reaction tank; a step of supplying the sample from a connector of the injection needle to the reaction tank; A dispensing method comprising: closing the valve and the gas inlet / outlet hole when it is detected that a sealing lid that seals a surface of the reaction tank into which the sample is injected is detected. 11. A step of dispensing a sample into a reaction tank for injecting the sample, which comprises a container-like member having a periphery sandwiched by a first bonding member and a second bonding member, and the reaction. In the tank, a sealing lid composed of a cover member sandwiched by a third bonding member that contacts the second bonding member and a fourth bonding member that faces the third bonding member is provided. A dispensing method comprising a step of fitting and a step of disposing the reaction tank in the mounting block. 12. The dispensing method according to claim 11, wherein the fitting step includes a step of bonding the second adhesive member and the third adhesive member with an adhesive. 13. The fitting step includes the step of fitting and fitting the concave portion and the convex portion provided in the second bonding member and the third bonding member, respectively. Item 11. The dispensing method according to Item 11.