JP2003166910A - Liquid-feeding mechanism and analyzer provided with the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an analyzer for easily and accurately performing various analyses, such as POC analysis and a compact and low-cost liquid-feeding mechanism suitable for feeding a liquid in the analyzer. <P>SOLUTION: The analyzer for biochemical measurement is constituted of a chip 80, a partition-deforming device, and an optical detecting device. The chip 80 comprises a plurality of liquid chambers 83 filled with liquid samples and reagent solutions, a capillary 84 connected to the liquid chambers 83 to make the liquids flow, and diaphragm membranes 85 for covering opening parts of the liquid chambers 83. By pressing the diaphragm membrane 85 using a plunger 90 of the partition deforming device, in such a way as to be deformed and protruded toward inside the liquid chamber 83, the capacity of the liquid chamber 83 is changed to feed the liquid in the liquid chamber 83 to the capillary 84 and analyze specific components in the liquid. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to the analysis of a trace amount sample,
The present invention relates to an analyzer that can easily perform detection. Further, the present invention relates to a liquid feeding mechanism suitable for liquid feeding in the analyzer.

[0002]

2. Description of the Related Art Analysis for bedside diagnosis (POC (point of care)) in which measurement necessary for medical diagnosis is performed near the patient.
(Analysis) and analysis of harmful substances in rivers and wastes at sites such as rivers and waste treatment plants (POU (point of use)
Analysis and measurement such as contamination inspection at each site of cooking, harvesting, and importing food, etc., where analysis / measurement is required at or near the site where analysis / measurement is required (hereinafter referred to as "POC analysis, etc.") The importance of (i.e.) is attracting attention, and in recent years, the development of detection methods and devices applied to such POC analysis has been emphasized. Further, such POC analysis and the like are required to be performed easily in a short time and at low cost.

Conventionally, for microanalysis, a GC-MS apparatus or an LC-MS apparatus is widely used in which a sample is separated by capillary gas chromatography (CGC), capillary liquid chromatography (CLC) or the like and then quantified by a mass spectrometer. Has been used. However, these mass spectrometers are suitable for use on site such as the bedside of patients, contaminated rivers, waste treatment plants, etc. due to the large mass spectrometer and complicated operation. Not not. Further, in an analyzer for medical diagnosis using blood or the like as a sample, it is desirable to dispose the portion in contact with the sample.

In order to solve these problems, therefore, a conventionally used analyzer is downsized, and a micro-TAS (micro total analysis) in which an extremely small amount of liquid reagent is reacted.
The study of applying the technology of system) to POC analysis has been advanced. In μTAS, a groove is formed on the surface of a glass or silicon chip of about 10 cm to several cm square and a reagent solution or sample is poured into the groove in order to make the amount of sample not only blood but a minute amount. A minute amount of sample is analyzed by performing separation and reaction (JP-A-2-245655, JP-A-3-226666, and JP-A-8-233778).
Publication, Analytical Chem. 69, 2626-2630 (1997) Ac
lara Biosciences etc.). This technique has the advantage that the amount of sample, the amount of reagent necessary for detection, the amount of waste such as consumables used for detection, and the amount of waste liquid are all small, and that the time required for detection is generally short. is there.

The applicant of the present invention also filed Japanese Patent Application No. 10-181586.
Specification (“mixing analyzer and mixing analysis method”), JP 2000-2675 A (“Capillary photothermal conversion analyzer”), JP 2000-2677 A (“analyzer”), International Publication WO99 / Patent applications have been filed for μTAS-related inventions such as Japanese Patent Application No. 64846 and Japanese Patent Application No. 11-227624 (“Analysis cartridge and liquid feeding control device”). In these publications or application specifications, it is described that a resin-made microchip is used as a chip, a thermal lens detection method is used as a detection method of a trace amount component, and the like.

The thermal lens detection method is a photothermal conversion detection method in which a sample in a liquid is excited by excitation light to form a so-called thermal lens, and the change of the thermal lens is measured by the detection light. Known from before (Japanese Patent Laid-Open No. 60-174)
933, A. C. Boccara et. a
l. , Appl. Phys. Lett. 36, 130,
1980, J. Liquid Chromatography 12, 2575-2585 (1
989), JP-A-10-142177 and JP-A-4-3.
No. 69467, Bunseki No. 4,280-28
4, 1997, M.A. Harada, et. al. , An
al. Chem. Vol. 65, 2938-2940,
1993, Kawanishi, et al. Proceedings of the 44th Annual Meeting of the Analytical Chemistry Society of Japan, p119, 1995, etc.).

As a method for measuring the components in the capillaries, a fluorescence method, an absorbance method and the like can be used in addition to the thermal lens detection method, but a high sensitivity is realized without an operation such as introduction of a fluorescent labeling substance. The thermal lens detection method is suitable because it is possible. On the other hand, as a technique for moving the liquid in the chip, that is, a liquid feeding method, a liquid feeding pump or a suction pump outside the chip is generally used to feed the liquid in the capillary of the chip (for example, S . Shoji,
et. al. , Sensors & ActuatorsB
8, 205-208, 1994, Bunseki No. 4, 2
80-284, 1997, M.I. Harada, et. a
l. , Anal. Chem. Vol. 65,2938-
2940, 1993, Kawanishi, et al. The Analytical Chemistry Society of Japan 44th
Annual Meeting Abstracts, p119, 1995, etc.). Also, connect the liquid reservoir outside the chip to the end of the capillary of the chip,
It has also been reported that the connection part is filled with a liquid, and liquid is fed into the chip by suction force using a siphon phenomenon (Sensor / Actuator / Week '97 General Symposium, Microsensor, Session 3 Abstracts 19-23).
Page "Toward the integration of chemistry" Takehiko Kitamori, Tsuguro Sawada Organized by: Next Generation Sensor Conference, 1997).

However, these methods, especially the method using the external pump, have the responsiveness of control, continuous change,
There are problems in terms of durability and quietness, which are important in medical practice. Further, since the liquid feed pump and the suction pump are used, the entire apparatus may be large, and leakage may occur at the connecting portion between the tip and the external pump. Further, in an apparatus that requires circulation of the liquid with the outside, such as an external pump or a device using a siphon phenomenon, it is necessary to provide a waste liquid reservoir, a reagent solution reservoir, a buffer liquid reservoir, etc. outside the chip, and to replenish the liquid. , Disposal, cleaning, etc., requires maintenance of the liquid reservoir.

[0009] These things greatly impair the convenience in POC analysis and the like. Further, when the siphon phenomenon due to the liquid reservoir outside the chip is used, in addition to discarding the stored liquid, labor such as deaeration of the tube connecting the chip and the external liquid reservoir in order to establish the siphon,
It will occur every time the chip is replaced. On the other hand, capillary electrophoresis itself or a method of sending a liquid by applying a voltage using an electroosmotic flow in a chip has been proposed (International Publication WO96 / 04547, S.M.
C. Jakobson, et. al. , Anal. Ch
em. Vol. 66, 4127-4132, 1994,
J. Liquid Chromatography 12, 2575-2585 (1989), JP-A-10-142177, JP-A-4-369467
Issue Bulletin).

However, both electrophoresis and electroosmotic flow
Since a voltage is applied to the liquid in the chip via the electrodes, the reagent composition or the sample composition may change due to electrolysis of the measurement reagent or the measurement sample on the electrode surface. In addition, a reagent or an electrolysis product of a sample may adhere to the inner surface of the capillary to change the zeta potential on the surface of the capillary, resulting in a phenomenon in which the liquid transfer rate changes. In addition, a freeze-dried solid reagent is placed in the cartridge, the blood sample is diluted with the dissolution diluent enclosed in the cartridge, and the solid reagent is dissolved with the diluted sample solution to perform an analytical reaction and quantify. A method is disclosed (Japanese Patent Publication No. 10-501340, Japanese Patent Publication No. 9-50473).
No. 2, etc.).

In this method, since the liquid is fed by centrifugal force, the liquid feeding direction is always the direction in which the centrifugal force acts, that is, the direction outward from the center of the circle of the circular cartridge. The solid reagent is placed in a small chamber along the circumference, which is located at the end of the flow path in the cartridge.The diluted sample flows into each small chamber, dissolves the solid reagent, reacts, and changes into absorbance. Is coming. However, due to the structure of the cartridge, the solid reagent is located at the final point of the flow path, so only one-step detection reaction with one reagent can be performed, and depending on the detection items, it may depend on the detection center or clinical laboratory of a hospital. It is unavoidable to adopt a reaction and reagent composition different from the detection reaction by the recommended method established by academic societies and government offices, which is carried out in Japan. Therefore, the correlation with the conventional inspection data may be low. Furthermore, depending on the test item, it may be difficult to perform analysis using the reaction mode of such a circular cartridge.

Further, as another method of feeding liquid by centrifugal force, a valve for closing the flow passage by using surface tension generated in the narrow groove or a valve for closing the flow passage by using wax is provided in the middle of the flow passage. There is also known a technology for providing and controlling liquid transfer (International Publication WO98 / 53311, International Publication WO).
97/21090 publication). However, a valve utilizing surface tension has a problem that it is difficult to control the liquid transfer because the valve is easily opened with a reagent solution containing a surfactant. In addition, a valve using wax has problems that the flow path becomes thicker, a heating mechanism for melting the wax is required, and components in the liquid are adsorbed by the wax. Further, these valves have a problem that once they are opened, they cannot be closed again.

Furthermore, a method of moving plasma inside the chip by an intake / exhaust pump outside the chip has been disclosed (Japanese Patent Laid-Open No. 9-196920 (immunity item), Japanese Patent Laid-Open No. 8-14539 (Biochemical item). ), JP-A-9-
196739 (dissolution liquid tip detection), JP-A-9-1
38195 gazette (analysis by measuring transmitted light of a porous material) and the like). In this method, the plasma itself is dissolved in the adhering reagent and simultaneously reacted, but as the main liquid feeding method, vacuum suction / pressurization from the end of the flow channel is adopted. Further, since the liquid supply inside the chip is adjusted via air, it is not necessary to exchange the liquid for liquid supply with the outside of the chip.

However, there is a problem in that an intake / exhaust pump for feeding liquid is required outside the chip, and an interface between the chip and the intake / exhaust pump (a leak-free connection structure) is required. On the other hand, a liquid feeding method by air bubbles in a closed capsule has been reported. This includes
There are liquid transfer due to thermal expansion of bubbles and liquid transfer due to generation of electrolytic gas. The former is performed with a closed chamber and a heating material and a filling liquid. The expansion of the heated bubbles generates a high pressure, which enables a quick liquid transfer. For example Naruse Y
According to the method of oshihiro et al., a light absorbing exothermic material is enclosed in a chamber filled with a liquid, and when the laser light is guided into the chamber by a glass fiber, the heat generated by the light absorbing exothermic material causes the bubbles in the chamber to expand and send. Liquid is carried out (US Pat. No. 5,210,817).

On the other hand, the latter is performed by a sealed chamber, an electrolytic solution filled in the chamber, and a pair of electrodes inserted in the chamber. When a voltage is applied to the electrodes, pressure is generated by the generation of electrolytic gas, and liquid is sent. For example, DAHopkins, Jr (USP567
1905) and CR Neagu et al. (CR Neagu, JG
E.Gardeniers, M. Elwenspoek, JJKelly, Journal of
Micro Electro Mechanical Systems, 5,1, 2-9, (199
6)) has reported some.

However, since the bubbles are compressible and elastic, the pressure loss is large when the liquid is sent into the narrow capillary channel. Therefore, in any of the methods, a long time is required from the beginning of the liquid flow to the stabilization. In some cases, the liquid vibrates in the flow direction,
There is a possibility that a problem may occur in that the liquid transfer is not stable at all. Furthermore, a method of pushing the filling liquid from the outside to send the liquid without using bubbles has been proposed, for example, a liquid sending by a piezoelectric element can be mentioned. A piezoelectric element can generate a large force with a relatively small amount of energy.

However, if the piezoelectric element is composed of a single crystal, the liquid can be moved only by a very small distance. In order to increase the stroke, a piezoelectric element is usually composed of a plurality of crystals, but then many components are required, which eventually increases the cost. Further, since the piezoelectric element is driven by a small current but requires a high voltage, it cannot be said that it is necessarily adapted to today's semiconductor circuits. Further, it is necessary to stack materials having different expansion coefficients to form a piezoelectric element, and moreover, accurate clearance is required in stacking, and thus it is difficult to miniaturize the piezoelectric element. Furthermore, since it is a reciprocating motion due to vibration, in order to convert it into a unidirectional force suitable for liquid transfer, it is necessary to use a plurality of valves having a check valve function, or to provide an electrical phase difference for a plurality of pumps. Since control is required, there is a problem that the entire system becomes very complicated. It is possible to perform liquid transfer by installing a module having a rectifying effect such as a diffuser in the flow path, but due to its structural characteristics, the rectifying effect cannot be expected unless the flow velocity is high. When the flow velocity is low, it is possible to increase the speed by narrowing the width of the flow channel, but this method increases the pressure loss in the flow channel, and it is necessary to improve the chip manufacturing accuracy and flow rate control accuracy. However, it becomes difficult to build a practical system because of the high cost.

Furthermore, liquid transfer based on the principle of attraction of electrostatic charge repulsion and opposite charge has also been reported. The liquid feeding force is proportional to the surface area of the electrostatic liquid feeding element, that is, the charging plate, and the applied voltage, and is inversely proportional to the square of the gap distance. Therefore, in electrostatic liquid transfer, it is possible to quickly transfer liquid with the minimum power. For example, in the method of R. Zengerle et al., The liquid in the chamber is pushed out by the electrostatic repulsion between the thin film electrode and the chip fixed electrode (R. Zengerle, A. Richter, H. Sandm.
aier, Micro Electro Mechanical Systems'92, 4, 19,
(1992)).

However, since the gap distance is sensitive to the liquid feeding, the actual liquid feeding is limited to a few μm. This narrow gap is sensitive to contamination by dust and the like, and easily attracts dust at a high electric field, and such contamination hinders proper liquid transfer. Further, since it does not require a large current but a high voltage, it cannot be said that it is necessarily applied to today's semiconductor circuits. Furthermore, a large-capacity charging plate is required to obtain a large liquid feeding force. Furthermore, since it is a reciprocating motion due to vibration, a plurality of pilot valves that are electrically controlled or have a check valve function are required to convert into a unidirectional force suitable for liquid transfer. There was a problem that the whole thing became very complicated.

[0020]

As described above, although there are many proposals as a liquid feeding technology to be provided to the device for performing the POC analysis and the like, the device aimed at by us is multi-item, small size, simple, short time and low cost. No one has yet been proposed that meets all of the requirements of. Specifically, the device is small and can be easily manufactured, the operation is not complicated, and the measurement can be easily performed.
A reagent cartridge that does not cause contamination and can be disposable, a test result that correlates with a conventional test method, a method that can measure concentration by a thermal lens method, etc., and a disposable cartridge that does not have an external pump There is a demand for a liquid-feeding mechanism capable of satisfying many requirements such as one having no connecting portion and one capable of performing a concentration standard solution reaction to ensure accuracy.

Therefore, the present invention solves the problems of the prior art as described above, has a simple mechanism, is small in size and low in cost, and performs various analyzes including POC analysis. It is an object of the present invention to provide a suitable liquid feeding mechanism. Another object of the present invention is to provide an analyzer that includes the liquid feeding mechanism and can perform various analyzes such as POC analysis easily and accurately in a short time.

[0022]

In order to solve the above-mentioned problems, the present invention has the following constitution. That is, in the liquid feeding mechanism according to the first aspect of the present invention, the liquid filled in the liquid tank is connected to the liquid tank by changing the volume of the liquid tank formed surrounded by the wall body. A liquid sending mechanism that sends the liquid stored in the flow path or another liquid tank connected to the flow path to the liquid tank, wherein at least a part of the wall is It is characterized in that it is constituted by a partition wall having elasticity that is deformable so as to project toward the inside or the outside of the liquid tank.

Further, a liquid sending mechanism according to a second aspect of the present invention is the liquid sending mechanism according to the first aspect, characterized by including partition wall deforming means for deforming by pressing or pulling the partition wall. . Further, the liquid feeding mechanism according to claim 3 of the present invention is the liquid feeding mechanism according to claim 2, wherein the partition wall deforming means includes a plunger that presses or pulls the partition wall while linearly moving. And

Further, the liquid feeding mechanism according to a fourth aspect of the present invention is the liquid feeding mechanism according to the third aspect, wherein the partition wall deforming means accommodates the plunger and guides the linear movement of the plunger. A cylinder, a driving means for linearly moving the plunger, a coupler arranged at an end portion of the cylinder for enclosing and enclosing the partition wall from the outside of the liquid tank, and a flow of gas in and out of a portion surrounded by the coupler. And a valve for controlling.

Further, a liquid sending mechanism according to a fifth aspect of the present invention is characterized in that, in the liquid sending mechanism according to the second aspect, the partition deforming means includes a roller for pressing the partition. With such a configuration, since no joint or connection structure is provided unlike the case of liquid transfer by a normal pump, accurate liquid transfer can be performed even with a small amount of liquid. Further, since the liquid inside the liquid tank does not come into direct contact with the external plunger or roller, even if the liquid tank portion is configured as a disposable cartridge, contamination does not occur. That is, since the sample that has been analyzed previously does not essentially remain, it is particularly suitable for an analyzer for medical diagnosis.

Further, a liquid sending mechanism according to a sixth aspect of the present invention is characterized in that, in the liquid sending mechanism according to any of the second to fifth aspects, a protrusion is provided on an outer surface of the partition wall.
If the partition wall is pressed, the accuracy of the liquid delivery amount can be determined not by the accuracy of the machine but by the size of the protrusion that contacts the tip of the plunger or the roller surface. This is advantageous in reducing the manufacturing cost and size of the liquid feeding mechanism. In addition, even if a protrusion is provided on the tip of the plunger or the roller surface and the outer surface of the flat partition wall without protrusions is pressed,
The same effect as described above can be obtained.

Further, in the case of pulling the partition wall, since the partition wall can be pulled by grasping the projection, the operation of pulling the partition wall is easy. The method of providing the protrusions on the outer surface of the partition is not particularly limited, and examples thereof include a method of forming by screen printing and a method of fixing a separately manufactured resin to the partition. Furthermore, a liquid sending mechanism according to a seventh aspect of the present invention is characterized in that, in the liquid sending mechanism according to any one of the first to sixth aspects, the partition wall is made of a material through which gas permeates and liquid does not permeate. And

Further, the liquid feeding mechanism according to claim 8 of the present invention is the liquid feeding mechanism according to claim 7, wherein the material is
It is characterized by being a porous membrane having hydrophobicity. Further, a liquid sending mechanism according to a ninth aspect of the present invention is the liquid sending mechanism according to the eighth aspect, characterized in that the porous membrane is formed of an organic polymer. With such a configuration, when the liquid tank or flow path is filled with the liquid, it acts as an air vent, and when the liquid is sent, it can be deformed to push or pull the liquid. It is possible to easily configure the liquid feeding mechanism without using the or mechanism.
Particularly, the configurations of claims 8 and 9 are preferable.

Furthermore, in the analyzer of the tenth aspect of the present invention, the liquid sample, or a mixed liquid of the liquid sample and the liquid reagent is caused to flow in the flow channel to obtain the sample or the mixed liquid. An analyzer for analyzing a predetermined component therein, comprising: a chip having a liquid tank filled with the sample or the mixed liquid; and the flow path connected to the liquid tank,
The liquid feeding mechanism according to any one of 1. Furthermore, the analyzer of claim 11 according to the present invention is
The analysis device according to claim 10, wherein the chip is configured by laminating a pair of flat plate-like members, and at least one of the pair of flat plate-like members has a groove on a plate surface, and the groove is provided. The flow path is formed by laminating the plate surfaces inside.

Furthermore, an analyzer according to claim 12 of the present invention is the analyzer according to claim 10 or 11, wherein:
The chip is provided with a liquid tank in which a dried reagent is stored, and the liquid reagent is charged in the liquid tank and the reagent is dissolved by the dissolution liquid, whereby the liquid reagent is stored in the chip. It is possible to prepare by. With such a configuration, it is not necessary to provide a special device or mechanism for preparing a reagent solution necessary for analysis outside the chip, and therefore the configuration of the analysis device can be simplified.

Further, an analyzer according to a thirteenth aspect of the present invention is an analyzer provided with a mechanism for simultaneously moving the liquids in a plurality of liquid tanks or a plurality of flow paths by using a liquid for liquid pushing. The analyzer according to any one of claims 10 to 12, wherein a plurality of liquid pushing channels are provided in which the liquid pushing liquid sent by the liquid sending mechanism according to any one of the claims 1 to 9 flows. Is connected to each of the liquid tanks or the plurality of flow paths.

With such a structure, operations such as mixing and diluting liquids necessary for analysis can be freely performed in the chip. Hereinafter, the liquid feeding mechanism and the analyzer of the present invention will be described in detail with reference to the drawings. [Chip] The chip included in the analyzer of the present invention includes the liquid tank filled with the liquid and the flow path through which the liquid is sent and the reaction of the minute amount is performed as described above. And
This flow path is preferably formed by laminating a pair of flat plate-shaped members having grooves on the plate surface. That is, as shown in FIG. 1, for example, a resin cover sheet 11 is attached to a plate surface 10b of the flat plate member 10 having the groove 10a on the plate surface 10b having the groove 10a with an adhesive or an adhesive tape. Then, the capillary 12 serving as a liquid flow path
A chip 1 having is formed.

This groove can be formed by a technique such as molding with a die or embossing. The shape and size of the groove are not particularly limited, but from the viewpoint of the current molding technology, the ratio of the width to the depth of the flow channel is 0.3 to 10.
It is preferable that the width and depth are each 0.5 μm or more, and from the viewpoint of the amount of sample and reagent required,
Each depth is preferably 500 μm or less.
When a resin is used as the material of the chip (flat plate member), it is required that the molding processability is good and that it is transparent when performing optical measurement. Preference is given to using. Although it is possible to use glass as the material of the chip, resin is preferable in view of cost.

Specifically, polystyrene, styrene resin such as styrene-acrylonitrile copolymer, methacrylic resin such as polymethylmethacrylate, methylmethacrylate-styrene copolymer, polycarbonate, polysulfone, polyethersulfone, polyetherimide,
Examples thereof include polyarylate, polymethylpentene, and 1,3-cyclohexadiene-based polymer. It is also possible to use these copolymers and blended products.

[Regarding Partition Wall and Liquid Delivery Mechanism] As shown in FIG. 2A, the liquid tank 14 is formed by a through hole provided in the flat plate member 10 and has elasticity so as to cover the opening thereof. A diaphragm film 15 (corresponding to a partition wall which is a constituent of the present invention) is attached. Since the diaphragm film 15 has flexibility and can be deformed,
When a pressing force is applied to the diaphragm film 15 from the outside of the chip 1 as shown in FIG.
5 is deformed and pushed into the liquid tank 14.

Then, since the volume of the liquid tank 14 is reduced by the deformation of the diaphragm film 15, the liquid is incompressible. Therefore, as shown by the arrow in FIG. The liquid L is pushed out to the capillary 12 by the volume change. Thereby, the liquid L in the liquid tank 14 can be moved. By performing liquid transfer using such a diaphragm mechanism, desired liquid transfer can be performed in the chip without using a complicated mechanism, equipment, etc. for connecting piping from the outside. Further, since it is not necessary to provide the chip with a dedicated facility such as a connection port for piping, the chip can be easily manufactured at low cost.

If the diaphragm film 15 is pulled and deformed so as to project to the outside of the liquid tank 14, the volume of the liquid tank 14 increases, and therefore, in contrast to the arrow in FIG. The liquid L in 12 is drawn into the liquid tank 14 by the volume change. This makes the capillary 1
The liquid in 2 or in another liquid tank connected to the capillary 12 can be moved. The diaphragm film 15 is not particularly limited as long as it is flexible and has a sheet shape that can be deformed by a small mechanism.
A (polytetrafluoroethylene) porous membrane is particularly preferred. P
If the diaphragm film 15 is formed of a film having a property that air is permeable but liquid is not permeable (which is repelled by water repellency) like the TFE porous film, the diaphragm film 15 has air permeability and the liquid tank 14 is When filled with a liquid, it becomes a functional film having water resistance. Therefore, when the liquid L is introduced from the capillary 12 into the empty liquid tank 14, the air in the liquid tank 14 is pushed out as it is filled with the liquid L and permeates the diaphragm film 15 to escape. Since the liquid L does not pass through due to the water repellency of 15, the liquid tank 14 can be completely filled with the liquid L.

In addition to PTFE, a porous film made of various materials can be used as the diaphragm film 15. However, it is preferable to use a hydrophobic organic polymer or an inorganic material so that the liquid does not pass through the membrane. The hydrophobic organic polymer preferably has a critical surface tension of about 0.04 N / m or less at 20 ° C., and examples thereof include polytetrafluoroethylene (PTFE), silicone, polyethylene, polypropylene, polystyrene and polyvinyl chloride. , Polycarbonate, polysulfone, polyether sulfone, polyarylate, polymethylpentene, 1,3-cyclohexadiene-based polymer and the like.

Although a cellulose acetate membrane can be used in some cases, in the case of a reagent solution to which a surfactant is added, a membrane having a strong hydrophobicity such as PTFE, silicone, polyethylene, etc. is more permeable to the liquid. It is preferable because it has a high water pressure resistance. Since the larger the water pressure resistance is, the higher the liquid pressure can be delivered, the higher the water pressure resistance is, the more preferable it is. It is preferably 0.1 kg / cm ² or more. Membranes having an average pore diameter of 0.1 μm to about 5 μm can be used, but considering that the smaller the pore diameter, the higher the water pressure resistance and the small amount of permeated air, the most preferable is about 0.1 μm. The film thickness is preferably 25 to 300 μm.

[Regarding Analytical Device] By utilizing such a principle, it is possible to form a branched channel in the chip 1 as shown in the schematic view of FIG. This chip 1 has a plurality of liquid tanks (A tank, D tank, S tank, W tank), and
The opening of the tank is covered with the diaphragm film 15 described above. Further, the D tank is also provided with a mechanism (not shown) for pressing and deforming the diaphragm film 15 to change the volume of the D tank, that is, a mechanism for feeding the liquid in the D tank.
The flow channel extending from the D tank is flow channel s at branch point d.
And the flow path a, and the flow path s communicates with the S tank and the flow path a communicates with the A tank. Further, the openings of the S tank and the A tank are covered with a film having both air permeability and water resistance, such as the above-mentioned PTFE film, and opened to atmospheric pressure.

For example, when the diaphragm film 15 of the D tank is pressed by a plunger and the diaphragm film 15 is deformed so as to project toward the inside of the D tank, the volume of the D tank decreases, so that the liquid in the D tank is reduced. Is pushed out of the D tank, reaches the branch point d through the flow path, and is branched into the flow path s and the flow path a. Since these flow paths are in the same closed space, the liquid is divided into the S tank and the A tank at a ratio according to the cross-sectional area of each flow path. The ratio of this divided flow rate is not determined by the flow rate or pressure generated by pressing, but is determined by the ratio of the cross-sectional area of each flow path because the S tank and A tank are open to the same atmospheric pressure and are constant. To be done.

For example, suppose that the S sample tank is filled with the sample liquid to be analyzed, the A tank is filled with the reaction reagent solution, and the upstream channel and the D tank are filled with the buffer solution functioning as the liquid for pushing.
The buffer solution is extruded by changing the volume of the tank and is sent to the S tank and the A tank. Then, the sample in the S tank and the reagent in the A tank are both extruded at the same pressure with the volume ratio determined by the cross-sectional areas of the channel s and the channel a. The extruded sample and reagent merge and mix at the merge point m, and are sent to the W tank which is a waste liquid reservoir.

In the above example, the diaphragm membrane 15 covering the D tank is pressed to feed the liquid, but instead of this, the diaphragm membrane covering the W tank is pulled to feed the liquid. Good. In this way, for example, the liquid feeding and mixing of the sample and the reagent required in the general biochemical measurement process can be realized by providing only the simple diaphragm structure and the capillary channel on the chip. Such a liquid delivery system is suitable as a basic element used in a small-sized and inexpensive analyzer for performing POC analysis and the like.

Further, when the liquid is sent from the D tank to only the S tank and filled, the following procedure may be performed. That is,
The openings of D tank, S tank, A tank, and W tank are covered with a PTFE porous membrane (diaphragm film) having flexibility, air permeability, and water resistance, respectively. Make the inside air permeate to the outside, and PT of A tank and W tank
The FE porous membrane is closed so that the air in both tanks does not permeate to the outside. Then, when the PTFE porous membrane of the D tank is pressed and deformed to change the volume of the D tank, the liquid in the D tank is pushed out of the D tank and is not split at the branch point d to only the flow path s. Liquid is delivered to the S tank.

Thus, a simple diaphragm structure,
If a mechanism for controlling the permeation of gas to the outside through the diaphragm membrane is provided, the liquid can be evenly sent to all of the plurality of liquid tanks or only a part of the plurality of liquid tanks. This can be easily done. For example, biochemistry,
In analysis in immunology or the like, a method of determining the concentration of a measurement item is generally used by using a calibration curve (calibration line) prepared by measuring a standard solution having a known concentration in advance. In POC analysis, etc., a disposable chip is often used to prevent contamination and improve usability. However, when the chip is made of resin, etc., the cross-sectional area of the flow channel may vary due to its manufacturing accuracy. Therefore, the disposable chip may be a factor that deteriorates reproducibility.

However, the concentration standard solution can be calibrated by first measuring the concentration standard solution to create a calibration line and then measuring the actual sample, so that the error in the liquid transfer amount caused by the accuracy of the flow path can be calibrated. Therefore, if the actual sample is measured twice, an accurate value calibrated for each individual chip can be obtained. Therefore, as described above, the present liquid supply system capable of performing desired liquid supply to a plurality of liquid tanks can use the same flow path for the concentration standard liquid and the actual sample, and It is possible to build a measurement system that is not affected by manufacturing accuracy.

[0047]

BEST MODE FOR CARRYING OUT THE INVENTION Embodiments of a liquid feeding mechanism and an analyzer equipped with the liquid feeding mechanism according to the present invention will be described in detail with reference to the drawings. The present embodiment shows an example of the present invention, and the present invention is not limited to this embodiment. FIG. 4 shows a schematic diagram for explaining the configuration of the analyzer for biochemical measurement. This analyzer consists of 8 liquid tanks (D tank, A tank, B tank, S tank, P tank, Q tank, R tank, W tank).
4) having a tank) and a flow path communicating with the liquid tank
It has 0. The chip 40 is formed by laminating a cover sheet on a flat plate-shaped member having a through hole and a groove. By laminating the cover sheet on the plate surface of the flat plate member having the groove, a capillary serving as a liquid flow path is formed, and a liquid tank is formed by the through hole.
The flat plate member and the cover sheet are made of a transparent thermoplastic resin.

The opening of each liquid tank is covered with a hydrophobic porous film 41 made of an organic polymer having elasticity. For example, in order to realize a very general biochemical measurement, a sample, a buffer solution for diluting the sample, and two kinds of reagent solutions are required. The P tank is a sample, the Q tank is a liquid tank that is filled with a buffer solution, and the R tank is a reservoir tank that stores the buffer solution that fills the entire flow path in advance.

Further, the S tank is a liquid tank into which a diluted sample in which the sample in the P tank and the buffer solution in the Q tank are mixed is introduced.
When the sample and the buffer solution are mixed, the hydrophobic porous membranes 41, 41 provided in the P tank and the Q tank may be pressed at the same time to be deformed to send the liquid. For the deformation of the hydrophobic porous film 41, a partition deformation device described later is used. Furthermore, tank A,
In the tank B, a dried reagent is stored in advance. Then, the buffer solution is supplied in the initial stage of filling the entire flow path with the buffer solution, and thereby the dry reagent is dissolved in the buffer solution. As a result, the tanks A and B become liquid tanks for filling the reagent solution. Become.

In the vicinity of the D tank, a partition deforming device having a plunger 43 that presses the hydrophobic porous membrane 41 of the D tank while linearly moving is arranged. When this partition deforming device is driven, the plunger 43 causes the hydrophobic porous membrane 41 of the D tank to move.
Is pressed to deform the hydrophobic porous film 41 so as to project toward the inside of the D tank. Then, since the volume of the D tank decreases, the buffer solution in the D tank is extruded and sent to the connecting flow path. As described above, since the buffer solution (functioning as a liquid for pushing liquid) is filled in the flow path in the initial stage, the diluted sample in the S tank and the reagent solutions in the A tank and the B tank are sequentially pushed out. Then, the liquid is sent to the connected flow paths s, a, and b.

The diluted sample and the reagent solution in the tank A are mixed in the flow path sa.
Then, the reagent solution in tank B is mixed and reacted in the mixing channel sab, and the degree of reaction can be measured by an optical method (for example, optical absorption rate by dye). Then, the mixed liquid is a liquid tank W which is a waste liquid reservoir.
Sent to. Since all the liquid after the reaction is sent to the liquid tank W, the liquid tank W needs to have a sufficient capacity. Further, even when the number of reagents used in the reaction is increased, liquid tanks (A ₁ , A ₂ , ...
By preparing a large number of A _n ), a reaction system of multiple reagents can be easily constructed by the same method as described above.

In the above example, the liquid was sent by pressing the hydrophobic porous membrane 41 covering the D tank with the plunger 43, but instead, the hydrophobic porous membrane covering the W tank. The liquid may be sent by pulling 41 with the plunger 43 and deforming it so as to project to the outside of the liquid tank. The flow channel configuration shown in FIG. 5 is an example,
The structure is not limited to the structure shown in FIG. 5 in which the other flow paths a1 to an are sequentially joined to the flow path s, and the structure such as a structure in which the flow path a1 and the flow path a2 are joined can be freely designed as desired.

Further, when it is necessary to retain a buffer solution necessary as a liquid for pushing a liquid or a liquid for dissolving a reagent in the chip for a long period of time, a storage container such as a blister pack or a potion pack is previously prepared. It is good to put it in and attach it to the tip. Further, if a mechanism is provided so that the storage container can be taken out at the time of analysis, the necessary reaction can be performed only by the chip without supplying a buffer solution from the outside of the analyzer. With such a configuration, the reaction can be performed without supplying a liquid such as a reagent from the outside, and the liquid after the reaction can be stored in the chip, so that it is not necessary to prepare a container for waste liquid, Therefore, it is effective for preventing contamination and making the analyzer free of maintenance.

Further, the dilution or extraction process of the sample specimen such as blood can be considered in the same manner, and most of the conceivable reagent reaction processes can be realized by using the above method. (Examples) The present invention will be described more specifically with reference to Examples. FIG. 6 is a conceptual diagram for explaining the overall configuration of all devices and equipment necessary for sample analysis, such as a chip, a liquid-feeding actuator unit (partition deforming device), and an optical detection device.

A sheet-type heater for controlling the temperature of the chip was mounted on the measuring table, and the chip having the above-described structure was mounted on the sheet-type heater. Then, the liquid feeding actuator unit was mounted on the chip. The liquid feeding actuator unit includes a plunger that presses the diaphragm film,
A drive device for driving the plunger and a mechanism for controlling permeation of gas from the liquid tank to the outside through the diaphragm membrane are provided. Then, the operation of the liquid feeding actuator unit is freely controlled by the central control CPU via the control circuit and the drive circuit to feed the liquid in the chip.

Further, in order to monitor the biochemical reaction process in the chip, a detecting device for detecting the concentration by the photothermal conversion method is provided. In this detection device, the optical measurement system is movable by a command from the overall control CPU, and optical measurement can be performed at any position on the chip plane. With this configuration,
No matter what the arrangement of the liquid tank (reagent liquid tank, sample liquid tank, etc.) or the flow path in the chip is, liquid transfer and measurement can be performed quickly.

FIG. 7 shows the flow path configuration of the chip 80 for analyzing biochemical measurement items (cholesterol, glucose, etc.) using a blood sample and a commonly used measurement reagent. The configuration of the chip 80 will be described later. The R tank is filled with the buffer solution in an amount capable of preliminarily filling the Q tank, D tank, A tank, B tank, and all the flow paths, and since the fluid pressure is generated, the entire buffer solution can be filled in the initial stage of the analysis. It is like this.

The P tank is filled with a blood sample, and the Q tank is filled with a buffer solution for diluting the blood sample. Then, the liquid feeding actuator unit is operated to feed the two liquids to the flow path p and the flow path q, respectively.
By merging at q, the S sample is filled with the blood sample diluted to a predetermined concentration. Further, the tank A and the tank B are pre-stored with the dried reagent, and when the buffer solution is introduced, the reagent is dissolved in the buffer solution, and the tank A and the tank B are filled with the desired reagent solution. It has become so.

FIG. 8 is a sectional view for explaining the structure of the chip 80. Groove 8 having a depth of 50 μm and a width of 100 to 500 μm
A rectangular flat plate-shaped member 1 having a plate surface 1a on one side
1 (thickness 2 mm, length 100 mm, width 150 mm)
It was manufactured by injection molding MMA (polymethylmethacrylate, acrylic resin). The material of the flat plate member 81 may be polycarbonate resin or glass. However,
In the case of glass, the groove 81a can be formed by etching or the like.

The liquid tank 83 is added to the flat plate member 81.
A through hole was formed by a drill. Then, a resin cover sheet 82 having a thickness of about 0.3 mm was attached to the plate surface of the flat plate-shaped member 81 having the groove 81a to obtain a chip 80. By attaching the cover sheet 82, the liquid L
A capillary 84 that serves as a flow path is formed. An ultraviolet curable resin 86 was used as an adhesive when the cover sheet 82 was attached. In consideration of performing optical measurement, it is preferable to use an adhesive made of a material whose optical performance such as light wavelength, refractive index, and transmittance is suitable for the measurement method. A system adhesive was used. In addition, you may stick using the double-sided adhesive tape which apply | coated the adhesive agent on the resin base material.

The opening of the liquid tank 83 is covered with a diaphragm film 85 (made of porous PTFE having a pore size of about 1 μm and a water resistance of 10 MPa) having flexibility, air permeability and water resistance. The diaphragm film 85 can be made of rubber, a vent film, a resin film, or the like, but it is necessary to have both the property of allowing air to pass through and the property of not allowing liquid to pass therethrough.
If it does not have such a property, air may be mixed into the liquid tank 83. If this is done, the pressure due to the deformation of the diaphragm film 85 will be consumed in the compression of the mixed air, so that accurate feeding is possible. There is a risk that the liquid will not be formed.

The diaphragm deforming mechanism (partition deforming means) in such a chip 80 will be described with reference to FIG. This diaphragm deformation mechanism includes a plunger 90 that linearly moves to press the diaphragm film 85, and a rotary motor 91 that is a power source that drives the plunger 90.
And a cylinder 93 provided with a ball screw 92 for converting the rotational movement of the rotary motor 91 into a linear movement of the plunger 90, accommodating the plunger 90 and guiding the movement of the plunger 90, and an end portion of the cylinder 93. And a coupler 94 that is integrally disposed in the liquid tank 83 to surround and seal the diaphragm film 85 from the outside of the liquid tank 83, and controls the inflow and outflow of gas in the portion surrounded by the coupler 94 to open the pressure of the portion to the atmospheric pressure. And a valve 95 for turning on and off.

The type of rotary motor 91 is not particularly limited, and a step motor or the like can be used. Regarding the motor performance, in the case of the present embodiment, it was sufficient if a thrust of 1N or more could be generated.
In addition, since the conversion mechanism using the ball screw 92 is provided, a force increasing effect and a displacement reducing effect can be expected, high control accuracy is not required, and if it is small, a commercially available product can be used without any problem. The diameter of the plunger 90 may be designed to be slightly smaller than the diameter of the opening of the liquid tank 83. Then, if the plunger 90 is the liquid tank 83,
Even if the liquid is set at a position deviated from the center position, the liquid transfer amount does not change due to the deformation of the diaphragm film 85, and accurate liquid transfer can be performed.

The chip 8 of the diaphragm deformation mechanism
It is desirable that the mounting to 0 is performed before the liquid is filled in the liquid tank 83. This is because if the liquid tank 83 is mounted after being filled with the liquid, the liquid inside may be inadvertently moved by the mounting operation. The graph of FIG. 10 shows the result of the liquid transfer in the chip 80 performed by such a diaphragm deformation mechanism. The horizontal axis of the graph shows the length of the portion of the plunger 90 inserted into the liquid tank 83 (insertion length), and the vertical axis shows the flow velocity of the liquid during liquid feeding. Further, the liquid tank 83 equipped with the diaphragm film 85.
Has a cylindrical shape and has a diameter of 1 mm (indicated by a triangle in the graph) and 3 mm (indicated by a circle in the graph).
The liquid was sent for Furthermore, the dotted line shown in the graph represents the theoretical value of the flow velocity of the liquid. The insertion speed of the plunger is 0.2 μm / s in all cases.

As can be seen from this graph, the liquid can be sent at a flow rate close to the theoretical value. Since the flow rate is determined by the cross-sectional area of the flow path, it is not necessary to have high accuracy in the flow rate itself.However, when controlling the reaction time, it may be necessary to control the flow rate. It is preferable that the accuracy is excellent. Further, it can be seen from this graph that the reaction time can be controlled by changing the diameter of the liquid tank on the liquid feeding side. In addition, on the outer surface of the diaphragm film 85 (the surface on the side where the diaphragm deformation mechanism is arranged),
It is preferable to form the protrusion 100 as shown in FIG. The protrusion 100 can be easily manufactured by screen printing using resin ink or the like.

If the size (particularly the height) of the protrusion 100 is set so as to cause a desired deformation when pressed by the plunger 90 as shown in FIG. It is possible to realize liquid transfer with a simpler mechanism without controlling the pushing size by the step motor as shown in FIG. This dimension setting can be easily controlled by changing the mask thickness or the like during screen printing. The structure shown in FIG. 11 (b) is a mechanism of the analyzer, in that a commercially available single-acting electromagnetic actuator can be used to produce a desired amount of liquid by simply pressing the diaphragm film 85. Has the advantage that it can be greatly simplified.

Further, as a simpler mechanism, there is a method of rolling the roller 101 on the tip 80 as shown in FIG. 11 (c). Roller 1 that can be manufactured at low cost
01, and the protrusion 100 of the diaphragm film 85.
Since a simple mechanism in which the roller 101 rolls so as to pass above is used, it is possible to further reduce the cost and positional accuracy of the diaphragm deformation mechanism. It is conceivable that various protrusions are present in the rolling portion of the roller 101, but this may be designed in advance so that it can be avoided.

For these, it is necessary to select the most appropriate method from the viewpoint of the ease of production, the production cost, the availability of the parts and materials, and the like. Next, referring to Table 1, a procedure for performing liquid transfer and analysis in the chip 80 of FIG. 7 using the diaphragm deformation mechanism shown in FIG. 9 will be described.

[0069]

[Table 1]

The eight liquid tanks of the chip 80 of FIG. 7 (P tank,
Q tank, D tank, S tank, A tank, B tank, R tank, W tank)
The diaphragm deformation mechanism shown in FIG. 9 is attached to each of the five liquid tanks other than the tank, the tank A, and the tank B. The P tank is filled with plasma separated from the blood, and the A and B tanks have dried reaction reagents attached to the bottom of the tank. Further, the R tank is filled with a buffer solution (initial state). Table 1 shows the contents of the operation for performing the liquid feeding and the analysis on the chip 80 and the state of the liquid tank after each operation. "Sample" described in Table 1 means that the sample exists in the liquid tank, and "-" means that the liquid tank is empty. Further, "dry test A" means that the dry A reagent is stored in the liquid tank, "dry test B" means that the dry B reagent is stored in the liquid tank, "buffer solution" is the buffer solution, and "reagent A" is the A solution. Reagent solution, "Reagent B", means that the B reagent solution is present in the liquid bath. Further, “closed” indicates that the operation for closing the valve 95 of the diaphragm deformation mechanism is performed, and “open” indicates that the same operation for opening the valve 95 is performed. Furthermore, "pressing" indicates that the operation of pressing the diaphragm membrane 85 with the plunger 90 is performed, and "NA" indicates that no operation is performed on the tank.

First, when the operation shown in the operation 1 of Table 1 is applied to the chip 80 in the initial state, the buffer solution (liquid for pushing) for feeding the liquid is filled in the entire flow path in the chip 80. It That is, the diaphragm deformation mechanism arranged in the R tank pushes out the buffer solution filled in the R tank,
Since the tank valve 95 is closed, the flow path sa
It flows through b, sa, s, a, and b into the S tank, A tank, and B tank. When the S tank is full, it flows into the Q tank through the flow paths pq and q, but since the P tank is already filled with the sample,
It does not flow into the flow path p to the P tank. Similarly, in tanks A and B, when they are filled with the buffer solution, they flow out into tank D. When the D tank is full, the Q tank, the D tank, the S tank, the A tank, and the B tank whose valves 95 are opened are filled with the buffer solution.

At this time, since the dry A reagent and the dry B reagent stored in the tanks A and B are dissolved in the buffer solution,
Each reagent solution is prepared in the B tank. The concentration of the reagent solution can be set to a desired value by the volume of the liquid tank and the amount of dry reagent. By such an operation, the reagent can be dissolved and the buffer solution for feeding the liquid can be filled, and the state shown in the state 1 of Table 1 can be obtained. Next, when the operation shown in the operation 2 of Table 1 is performed, the sample is diluted. That is, when only the valve 95 of the W tank is opened and the sample and the buffer solution are simultaneously pushed out by the diaphragm deformation mechanism arranged in the P tank and the Q tank, both liquids flow into the S tank and the sample is stored in the S tank. A diluted sample is prepared by diluting with a buffer solution. The dilution ratio depends on the cross-sectional area of the flow path and the speed at which the diaphragm film 85 is deformed (the plunger 90
Insertion speed) can be controlled to a desired value. By such an operation, the S tank is filled with the diluted sample, and
The state may be as shown in state 2. As a result, the S tank was filled with the diluted sample, and the A tank and the B tank were filled with the respective reagent solutions, and the reaction was ready.

Finally, when the operation shown in the operation 3 of Table 1 is performed, the diluted sample is extruded from the S tank and the reagent solutions are extruded from the A tank and the B tank, respectively, and flow through the flow paths s, a and b. To W tank. That is, the buffer solution filled in the D tank is pushed out by the diaphragm deformation mechanism arranged in the D tank, and the pressure is evenly transmitted to the S tank, the A tank, and the B tank through the three-branch flow path. As a result, the pressure is evenly transmitted to the channels s and a, and the liquid (diluted sample and A reagent solution) flows at a flow rate corresponding to the cross-sectional areas of the channels s and a.

Both liquids flow into the flow path sa and are mixed to form the flow path s.
At a, the reaction between the sample and the A reagent occurs. The mixed liquid flows toward the confluence with the flow path b while reacting, and the reaction time can be controlled by the flow velocity and the length of the flow path sa, when the reaction time is the time of flowing through the flow path sa. After the reaction between the sample and the reagent A proceeds for a predetermined time, the mixture reaches the mixing point with the flow path b. As a result, the reaction is further caused by mixing with the B reagent, so that the mixed solution capable of performing the final concentration measurement flows into the flow path sab. After mixing
After a reaction time suitable for the reagent reaction system, the dye concentration is labeled. If the length of the flow path sab is set so that a sufficient reaction time can be taken in consideration of the flow velocity, immediately after the reaction, the inside of the flow path is passed through the resin substrate using the optical measuring device as described in FIG. It is possible to measure the concentration of the solution.

The mixed solution whose concentration has been measured reaches the W tank (waste liquid tank) and is stored as a waste liquid. As described above, since the chip 80 after the concentration measurement can be disposed as it is, it is possible to construct an analysis system in which an analysis operator is not infected with a sample.
The mixing ratio may be adjusted for each reagent reaction system at the confluence of the flow paths, but in this method, the mixing ratio can be adjusted by the liquid flow rate ratio. The flow rate ratio of the flow paths s and a is determined by the cross-sectional area ratio as described above. In the usual case, the groove depth is often constant because of the ease of forming a mother die when molding the chip 80, but the cross-sectional area of the flow channel can be adjusted by changing the groove width. .

Therefore, when it is desired to set the mixing ratio of the sample and the reagent to, for example, 1: 2, the flow channel width ratio is designed to be 1: 2, and the diaphragm film 85 is simply pressed to deliver the liquid. , A mixing ratio of 1: 2 is obtained. Of course, the cross-sectional area may be adjusted by changing the depth of the channel or both the depth and the width of the channel. Furthermore, the reaction time can also be controlled by controlling the flow rate (flow velocity) by an external mechanism. Since the flow velocity is the flow rate divided by the cross-sectional area, in the above example, 120 after mixing
Since a reaction time of seconds is required, the flow rate is 100 μm / s
(Cross section area 0.005 mm ² , flow rate 500 nL / sec)
As a result, the reaction time was secured by designing a reaction channel of 12 mm. Also, instead of operating continuously,
When the deformation of the diaphragm film 85 is controlled to stop the flow of the liquid, the reaction time becomes infinite, and the degree of freedom in design increases.

Further, in this method, even if there is a dimensional difference in the flow path, calibration can be performed by using the concentration standard solution. Prepare a concentration standard solution with a known concentration in advance, and once execute the same reaction process as the actual reaction to create a calibration curve. This calibration curve gives the correspondence between the concentration and the signal at the chip, which is how much signal output is obtained for a known concentration in advance. Therefore, if a calibration curve is created for each chip before actual measurement of a sample, it is possible to obtain an analysis result that excludes fluctuations in concentration due to the dimensions of the flow channel and the amount of reagent.

Further, according to the present method, since the liquid is sent in the completely closed flow path, the disturbance factors such as bubbles are less likely to occur,
In addition, even if it occurs for some reason, the plunger 9
Since it has a function of forcibly washing off at 0, it is unlikely to be a disturbance factor given to the measurement system. Next, the result of actual analysis of the sample will be described. As a sample, standard serum (for lipid measurement, determiner, manufactured by Kyowa Medix Co., Ltd.) was used and diluted with Good buffer (50 mM, pH 7.0, manufactured by Dojindo Laboratories) to give a cholesterol concentration of 0.5.
What was adjusted to 0,100 mg / dL was used as a sample. Further, HA test Wako (manufactured by Wako Pure Chemical Industries, Ltd.), which is a reagent kit for cholesterol measurement, was used as the reagent solution, and the enzyme solution A and the enzyme solution B were sealed in tanks A and B of the chip of FIG. 7, respectively. A PTFE film was used as the diaphragm film 85, and liquid was sent by pressing the PTFE film with the plunger 90.

As the detecting device, a lock-in type thermal lens measuring device (NA of objective lens: 0.4) with a frequency of 1 kHz, which was uniquely constructed, was used. Then, the thermal lens signal was measured using a laser beam having a wavelength of 630 nm and an irradiation position intensity of 2.8 mW as the excitation light and using a laser beam having a wavelength of 780 nm and an irradiation position intensity of 3.2 mW as the probe light. Further, the flow path of the used chip has a depth of 5
The volume of the liquid tank for the sample is 2 μL, and the volume of the liquid tank for the reagent solution is 2 μL.

The analysis result is shown in the graph of FIG. As can be seen from the graph, there was a high correlation between the cholesterol concentration in the adjusted diluted sample and the output of the thermal lens signal obtained by the thermal lens measurement. From this result, it became clear that biochemical measurement can be performed by the chip.

[0081]

As described above, the analyzer of the present invention is
Various analyzes including OC analysis can be performed easily and accurately. Further, the liquid feeding mechanism of the present invention is suitable for liquid feeding in the analyzer, and is small in size and low in cost.

[Brief description of drawings]

FIG. 1 is a cross-sectional view illustrating the configuration of a chip of an analyzer according to the present invention.

FIG. 2 is a cross-sectional view illustrating a diaphragm mechanism adopted in the liquid feeding mechanism of the present invention.

FIG. 3 is a schematic diagram illustrating a channel configuration of a chip of the analyzer of the present invention.

FIG. 4 is a schematic diagram illustrating the configuration of an embodiment of an analyzer of the present invention.

5 is a schematic diagram illustrating a configuration of a modified example of the analyzer of FIG.

FIG. 6 is a conceptual diagram illustrating the overall configuration of the analyzer of the present invention.

FIG. 7 is a schematic diagram illustrating a flow path configuration of a chip of an analyzer according to an example.

FIG. 8 is a cross-sectional view illustrating the configuration of the chip of the analyzer of the example.

FIG. 9 is a cross-sectional view illustrating the configuration of a diaphragm deformation mechanism of the analyzer of the example.

FIG. 10 is a graph showing the correlation between the insertion length of the plunger and the flow velocity of the liquid.

11 is a cross-sectional view illustrating the configuration of a modified example of the diaphragm deforming mechanism of FIG.

FIG. 12 is a graph showing the correlation between the cholesterol concentration in a sample and the output of a thermal lens signal.

[Explanation of symbols]

1,40,80 chips 10,81 Flat plate member 10a, 81a groove 10b Board surface 11,82 Cover sheet 12,84 capillaries 14,83 Liquid tank 15,85 diaphragm membrane L liquid 43,90 Plunger 91 rotary motor 93 cylinders 94 coupler 95 valves 100 protrusions 101 roller

Continued front page    F-term (reference) 2G042 CA10 CB03 FB02 FB10 HA02                       HA03 HA06 HA07                 2G052 AA28 AA30 AB16 AD06 AD26                       AD46 CA03 CA04 CA11 DA05                       DA08 DA09 DA12 DA22 FB03                       FB09 GA11 JA01 JA07 JA09                       JA16                 2G058 AA01 CA01 CC01 CC18 CC19                       DA01 DA03 DA07 EA03 EA05                       EA14 EB01 EB24 FA07 GA01                       GE02                 4G068 AA01 AB11 AD12 AD24 AD31

Claims (13)

[Claims] 1. A liquid filled in the liquid tank is sent to a flow path connected to the liquid tank by changing the volume of the liquid tank formed surrounded by a wall body, or the flow path. Alternatively, in a liquid sending mechanism that sends a liquid stored in another liquid tank connected to the flow path to the liquid tank, at least a part of the wall body projects toward the inside or the outside of the liquid tank. A liquid delivery mechanism characterized by being constituted by a partition wall having elasticity that can be deformed. 2. The liquid feeding mechanism according to claim 1, further comprising partition wall deforming means for deforming by pressing or pulling the partition wall. 3. The liquid delivery mechanism according to claim 2, wherein the partition deforming means includes a plunger that presses or pulls the partition while linearly moving. 4. The partition deformation means includes a cylinder that accommodates the plunger and guides the linear movement of the plunger, and a drive means that linearly moves the plunger.
A coupler provided at an end of the cylinder for enclosing and sealing the partition wall from the outside of the liquid tank, and a valve for controlling gas flow in and out of a portion surrounded by the coupler. The liquid feeding mechanism according to item 3. 5. The liquid delivery mechanism according to claim 2, wherein the partition deformation means includes a roller that presses the partition. 6. The liquid feeding mechanism according to claim 2, wherein a protrusion is provided on an outer surface of the partition wall. 7. The liquid feeding mechanism according to claim 1, wherein the partition wall is made of a material that is permeable to gas and impermeable to liquid. 8. The liquid transfer mechanism according to claim 7, wherein the material is a porous film having hydrophobicity. 9. The liquid sending mechanism according to claim 8, wherein the porous film is formed of an organic polymer. 10. An analyzer for analyzing a predetermined component in the sample or the mixed liquid by flowing a liquid sample or a mixed liquid of the liquid sample and the liquid reagent into a channel. A chip having a liquid tank filled with the sample or the mixed liquid and the flow path connected to the liquid tank, and the liquid sending mechanism according to claim 1. An analyzer characterized by the above. 11. The chip is constituted by laminating a pair of flat plate-like members, and at least one of the pair of flat plate-like members has a groove on a plate surface, and the plate surface having the groove is an inner side. The analyzer according to claim 10, wherein the flow path is formed by laminating the channels. 12. The chip is provided with a liquid tank in which a dried reagent is stored, and a solution is charged into the liquid tank and the reagent is dissolved by the solution to obtain the liquid state. The analysis device according to claim 10 or 11, wherein a reagent can be prepared in the chip. 13. An analyzer according to claim 1, further comprising a mechanism for simultaneously moving the liquids in the plurality of liquid tanks or the plurality of channels by the liquid for liquid pushing. 13. The liquid pushing flow path through which the liquid pushing liquid sent by a liquid mechanism flows is connected to the plurality of liquid tanks or the plurality of flow passages, respectively. The analyzer described.