JP2004226207A - 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 a 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, a liquid-feeding apparatus, and an optical detecting device. The chip comprises a plurality of liquid chambers filled with a liquid, a channel connected to the liquid chambers, and diaphragm membranes for covering opening parts of the liquid chambers. The liquid-feeding apparatus comprises a pressure generating means, a non-compressible medium, a sealed container, and a diaphragm member. Then, by pressing the diaphragm membrane for feeding a liquid being provided at the chip by using the liquid-feeding apparatus, and changing the capacity of the liquid chamber, an aimed amount of liquid in the liquid in the liquid chamber is fed to the channel. <P>COPYRIGHT: (C)2004,JPO&NCIPI

Description

[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to an analyzer that can easily analyze and detect a trace sample. The present invention also relates to a liquid sending mechanism suitable for sending a liquid in the analyzer.
[0002]
[Prior art]
Analysis for bedside diagnosis (POC (point of care) analysis) that performs measurements necessary for medical diagnosis in the vicinity of patients and analysis of harmful substances in rivers and wastes are performed at sites such as rivers and waste disposal sites. Performing analysis / measurement at or near the site where analysis / measurement is required, such as point-of-use (POU) analysis or contamination inspection at each site of food preparation, harvesting, and importing (Hereinafter, collectively referred to as “POC analysis etc.”) has attracted attention. In recent years, development of detection methods and devices applied to such POC analysis and the like has been gaining importance. It is required that such POC analysis and the like be performed simply, in a short time, and at low cost.
[0003]
Conventionally, a GC-MS device or an LC-MS device that separates a sample by capillary gas chromatography (CGC), capillary liquid chromatography (CLC), or the like, and then quantifies the separated mass spectrometer has been widely used for microanalysis. Was. However, due to the large size of the mass spectrometer and the complicated operation, these analyzers are suitable for use in on-site measurements such as on the bedside of patients, contaminated rivers, and near waste disposal sites. Not. Further, in an analyzer for medical diagnosis using blood or the like as a sample, it is desirable that a portion touched by the sample is disposable.
[0004]
In order to solve these problems, studies have been made to reduce the size of conventionally used analyzers and apply μTAS (micro total analysis system) technology for reacting a trace amount of liquid reagent to POC analysis and the like. did it. In μTAS, a groove is formed on the surface of a glass or silicon chip of about 10 cm to several cm square or less in order to minimize the amount of a sample, not just blood, and a reagent solution or a sample is allowed to flow through the groove. A small amount of sample is analyzed by performing separation and reaction (JP-A-2-245655, JP-A-3-226666, JP-A-8-233778, Analytical Chem. 69, 2626-2630 (1997). ) Aclara Biosciences). This technique has the advantage that the amount of specimen, the amount of reagent required for detection, the amount of waste such as consumables used for detection, and the amount of waste liquid are all reduced, and the time required for detection is generally short. is there.
[0005]
The applicant of the present application also discloses Japanese Patent Application No. 10-181586 (“mixing analysis device and mixing analysis method”), JP-A-2000-2675 (“capillary photothermal conversion analyzer”), and JP-A-2000-2677. ("Analyzer"), International Patent Application WO 99/64846, and Japanese Patent Application No. 11-227624 ("Analysis Cartridge and Liquid Feeding Controller") have filed applications for µTAS-related inventions.
[0006]
These publications or application specifications also describe the use of a resin microchip as a chip, and the use of a thermal lens detection method as a method for detecting a trace component.
The thermal lens detection method is a photothermal conversion detection method in which a sample in a liquid is excited with excitation light to form a so-called thermal lens, and the change in the thermal lens is measured with the detection light. (Japanese Patent Application Laid-Open No. Sho 60-174933, A.C. Bocchara et. Al., Appl. Phys. Lett. 36, 130, 1980, J. Liquid Chromatography 12, 2575-2585 (1989); 10-142177, JP-A-4-369467, Bunseki No. 4,280-284,1997, M. Harada, et.al., Anal.Chem.Vol.65, 2938-2940, 1993, Kawanishi , Et al., Proceedings of the 44th Annual Meeting of the Japan Society for Analytical Chemistry, p119, 1995).
[0007]
As a method for measuring the components in the capillary, a fluorescence method or an absorbance method can be used in addition to the thermal lens detection method, but since high sensitivity can be realized without performing an operation such as introduction of a fluorescent labeling substance, Thermal lens detection is suitable.
On the other hand, a method of feeding a liquid by applying a voltage using a capillary electrophoresis itself or an electroosmotic flow in a chip has also been proposed (International Publication WO96 / 044747, SC Jakobson, et. Al.). Chem., Vol.66, 4127-4132, 1994, J. Liquid Chromatography 12, 2575-2585 (1989), JP-A-10-142177, JP-A-4-369467, and the like.
[0008]
However, in both electrophoresis and electroosmotic flow, a voltage is applied to the liquid in the chip via the electrodes, which causes electrolysis of the measurement reagent or measurement sample on the electrode surface, which changes the reagent composition or sample composition. There is. In addition, a phenomenon may occur in which the electrolysis product of the reagent or the sample adheres to the inner surface of the capillary, changes the zeta potential on the surface of the capillary, and changes the liquid sending speed.
[0009]
In addition, a lyophilized solid reagent is placed in a cartridge, a blood sample is diluted with a dissolution diluent sealed in the cartridge, and the solid reagent is further dissolved in the diluted sample solution, and an analysis reaction is performed to quantify. Methods have been disclosed (Japanese Patent Publication No. 10-501340, Japanese Patent Publication No. 9-504732, etc.).
In this method, since the liquid is sent by centrifugal force, the liquid sending direction is always the direction in which the centrifugal force acts, that is, the direction going outward from the center of the circle of the circular cartridge. The solid reagent is placed in a small chamber along the circumference, 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 the absorbance. Is coming.
[0010]
However, due to the structure of the cartridge, the solid reagent is located at the final point of the flow path, so that only one-step detection reaction can be performed with one reagent. For example, a reaction and a reagent composition different from the detection reaction according to the recommended method defined by the academic society or the government office must be adopted. Therefore, the correlation with the conventional test data may be low. Further, depending on the inspection item, it may be difficult to perform analysis by such a reaction mode of the circular cartridge.
[0011]
On the other hand, there has been reported a liquid feeding method using air bubbles in a sealed capsule. This includes liquid sending by thermal expansion of bubbles and liquid sending by generation of electrolytic gas. The former is performed by a closed chamber, a heating material and a filling liquid. The expansion of the heated bubble generates a high pressure and enables a fast liquid transfer. For example, in the method of Naruse Yoshihiro et al., A light-absorbing heat-generating material is sealed in a chamber filled with a liquid, and laser light is guided into the chamber with glass fibers. In this case, the liquid is sent by the pressure switch (US Pat. No. 5,210,817).
[0012]
On the other hand, the latter is performed by using a sealed chamber, an electrolytic solution filled therein, and a pair of electrodes inserted into the chamber. A pressure is generated by the generation of an electrolytic gas by applying a voltage to the electrode, and the liquid is sent. For example, D. A. Hopkins, Jr (US Pat. No. 5,671,905) and C.I. R. Neagu et al. (Reported by CR Neagu, JGE Gardeniers, M. Elwenspoek, JJ Kelly, Journal of Micro Electro Mechanical Systems, 5, 1, 96). There is something.
[0013]
However, since bubbles have compressibility, when sending a liquid into a narrow capillary channel, a long time is required from the start of the flow of the liquid until the liquid is stabilized. In some cases, the liquid may vibrate in the flow direction, and a problem may occur in which the liquid is not completely stabilized.
Furthermore, a method has been proposed in which the filling liquid is pushed from the outside without using bubbles to send the liquid, and for example, liquid feeding by a piezoelectric element can be mentioned. A piezoelectric element can generate a large force with a relatively small amount of energy.
[0014]
However, when the piezoelectric element is formed of a single crystal, the liquid can be pushed only a very small distance. In order to increase the stroke, a piezoelectric element is usually formed of a plurality of crystals, but this requires many parts, which eventually increases the cost.
Further, the piezoelectric element is driven by a small current but requires a high voltage, and therefore cannot always be said to be suitable for today's semiconductor circuits. Further, it is necessary to form a piezoelectric element by laminating materials having different expansion coefficients, and since accurate lamination is required for lamination, it is difficult to reduce the size. Furthermore, since the reciprocating motion is caused by vibration, in order to convert the force into a unidirectional force suitable for liquid feeding, a plurality of valves having a check valve function are required, and an electrical pump for providing a phase difference to a plurality of pumps is required. Since control is required, there has been a problem that the entire system becomes very complicated. It is also possible to feed liquid by installing a module having a rectifying effect such as a diffuser in the flow path, but due to its structural characteristics, a rectifying effect cannot be expected unless the flow velocity is high. In the case of low flow velocity, it is possible to increase the speed by narrowing the flow path width, but this method increases the pressure loss in the flow path, and it is necessary to increase the chip manufacturing precision and flow rate control precision For this reason, it is difficult to construct a practical system for reasons such as high cost.
[0015]
Furthermore, liquid transfer based on the principle of electrostatic charge repulsion and counter charge inquiries has been reported. For example, R. In the method of 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. Sandmaier, Micro Electro Mechanical Systems '92, 4, 19, (1992)).
[0016]
However, the gap distance is sensitive to liquid sending, and is limited to a few μm in actual liquid sending. This narrow gap is susceptible to contamination by dust and the like, and it is easy to attract dust in a high electric field, and such contamination hinders appropriate liquid sending. Further, since a large current is not required but a high voltage is required, it cannot be said that the semiconductor circuit is necessarily adapted to today's semiconductor circuits. Further, a large-capacity charged plate is required to obtain a large liquid sending power. Furthermore, since the reciprocating motion is caused by vibration, a plurality of pilot valves that are electrically controlled or have a check valve function are required to convert the force into a unidirectional force suitable for liquid supply, and the system There was a problem that the whole was very complicated.
[0017]
[Patent Document 1]
U.S. Pat. No. 5,210,817
[Patent Document 2]
U.S. Pat. No. 5,671,905
[Non-patent document 1]
C. R. Neagu, J .; G. FIG. E. FIG. Gardeniers, M .; Elwenspoek, J .; J. Kelly, "Journal of Micro Electro Mechanical Systems", 1996, Vol. 5, No. 1, p. 2-9
[Non-patent document 2]
R. Zengerle, A .; Richter, H .; Sandmaier, "Micro Electro Mechanical Systems '92", 1992, Vol. 4, p. 19
[0018]
[Problems to be solved by the invention]
As described above, there are many proposals as a liquid transfer technique to be provided to an apparatus for performing POC analysis and the like, but none of the techniques that meet all of the demands of the equipment of various items, small size, simple, short time, and low cost are still available. Not proposed. Specifically, the device is small and easy to manufacture, the device can be easily measured without complicated operations, the analysis chip can be disposable from the viewpoint of preventing contamination, and the test results are the same as those of the conventional test method. Correlation, concentration measurement by photothermal conversion detection method (thermal lens method), etc., lack of complicated connection mechanism between external liquid sending device and analysis chip, concentration to ensure accuracy There is a demand for a liquid sending mechanism that can satisfy many necessary conditions, such as those capable of performing a standard solution reaction.
[0019]
Therefore, the present invention solves the problems of the prior art as described above, and its mechanism is simple, compact and low-cost, and is suitable for an analyzer that performs various analyzes including POC analysis. It is an object to provide a liquid feeding mechanism. It is another object of the present invention to provide an analyzer that includes the liquid feeding mechanism and can easily and accurately perform various analyzes including a POC analysis in a short time.
[0020]
[Means for Solving the Problems]
In order to solve the above problem, the present invention has the following configuration. That is, in the liquid feeding mechanism of claim 1 according to 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 by being surrounded by the wall. A liquid sending mechanism for sending to the flow path, or for sending the liquid contained in another liquid tank connected to the flow path or the flow path to the liquid tank, wherein at least a part of the wall body is A diaphragm having elasticity capable of being deformed so as to protrude toward the inside or outside of the liquid tank, and a diaphragm member in contact with an outer surface of the partition, and a diaphragm in contact with the diaphragm among the surfaces of the diaphragm member. A non-compressible medium in contact with the non-compressible medium, a container for storing the incompressible medium, and pressure generating means for applying pressure to the incompressible medium, wherein the incompressible medium includes the diaphragm member and the Container and Enclosed have been sealed in the closed space, is loaded on the partition walls the pressure through the incompressible medium and said diaphragm member, wherein said partition wall is adapted to be deformed.
[0021]
According to a second aspect of the present invention, in the liquid feeding mechanism according to the first aspect, the container and the pressure generating unit are connected by a tubular member filled with an incompressible medium. It is characterized.
Further, in the liquid feeding mechanism according to a third aspect of the present invention, in the liquid feeding mechanism according to the first or second aspect, the pressure generating means generates the pressure using a reciprocating positive displacement pump mechanism. It is characterized by making it.
[0022]
Further, in the liquid feeding mechanism according to a fourth aspect of the present invention, in the liquid feeding mechanism according to the first or second aspect, the pressure generating means generates the pressure using a rotary positive displacement pump mechanism. It is characterized by making it.
Further, in the liquid feeding mechanism according to a fifth aspect of the present invention, in the liquid feeding mechanism according to the first or second aspect, the pressure generating means uses the volume change accompanying a phase change of the substance to generate the pressure. Is generated.
[0023]
Further, according to a sixth aspect of the present invention, there is provided the liquid feeding mechanism according to the fifth aspect, wherein the heating device and the cooling device control the temperature of the substance to cause the substance to undergo a volume change due to a phase change. It is characterized by including at least one of the devices.
Further, in the liquid feeding mechanism according to claim 7 of the present invention, in the liquid feeding mechanism according to claim 1, the incompressible medium is formed of a material having rubber elasticity, and the pressure generating means is formed with a hole or a crack. , A non-compressible medium provided with a hole or crack that is continuous with a hole or crack in the container, and a pin that reciprocates by being inserted into the hole or crack of the container and the non-compressible medium. Or a thin section.
[0024]
Further, in the liquid feeding mechanism according to claim 8 of the present invention, in the liquid feeding mechanism according to any one of claims 1 to 7, the pressure in the vicinity of a contact portion between the partition and the diaphragm member is reduced to less than the atmospheric pressure. And a means for bringing the partition and the diaphragm member into close contact with each other and releasing the close contact between the partition and the diaphragm member by setting the vicinity of the contact portion to atmospheric pressure.
[0025]
Further, in the liquid feeding mechanism according to claim 9 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 by being surrounded by the wall. A liquid sending mechanism for sending to the flow path, or for sending the liquid contained in another liquid tank connected to the flow path or the flow path to the liquid tank, wherein at least a part of the wall body is A partition that has elasticity that can be deformed so as to protrude toward the inside or outside of the liquid tank, and is in contact with the outer surface of the partition, an incompressible medium, and a container that stores the incompressible medium, Pressure generating means for applying pressure to the incompressible medium, wherein the incompressible medium is sealed in a closed space surrounded by the partition and the container, and the pressure is the incompressible medium. Is applied to the partition via Characterized in that it adapted to form.
[0026]
Further, in the liquid sending mechanism according to claim 10 of the present invention, in the liquid sending mechanism according to claim 9, the container and the pressure generating means are connected by a tubular member filled with an incompressible medium. It is characterized.
Furthermore, in the liquid feeding mechanism according to claim 11 of the present invention, in the liquid feeding mechanism according to claim 9 or 10, the pressure generating means generates the pressure using a reciprocating positive displacement pump mechanism. It is characterized by making it.
[0027]
Further, in the liquid feeding mechanism according to a twelfth aspect of the present invention, in the liquid feeding mechanism according to the ninth or tenth aspect, the pressure generating means generates the pressure using a rotary positive displacement pump mechanism. It is characterized by making it.
Further, according to a thirteenth aspect of the present invention, in the liquid feeding mechanism according to the ninth or tenth aspect, the pressure generating means uses the volume change accompanying the phase change of the substance to generate the pressure. Is generated.
[0028]
Further, in the liquid sending mechanism according to claim 14 of the present invention, in the liquid sending mechanism according to claim 13, a heating device and a cooling device that control a temperature of the substance to cause a volume change in the substance due to a phase change. It is characterized by including at least one of the devices.
Further, in the liquid sending mechanism according to claim 15 of the present invention, in the liquid sending mechanism according to claim 9, the incompressible medium is made of a material having rubber elasticity, and the pressure generating means is provided with a hole or a crack. , A non-compressible medium provided with a hole or crack that is continuous with a hole or crack in the container, and a pin that reciprocates by being inserted into the hole or crack of the container and the non-compressible medium. Or a thin section.
[0029]
Furthermore, in the liquid sending mechanism according to claim 16 of the present invention, in the liquid sending mechanism according to any one of claims 9 to 15, at least a part of a contact portion between the partition and the incompressible medium is bonded. It is characterized by the following.
Further, the liquid sending mechanism according to claim 17 of the present invention is characterized in that, in the liquid sending mechanism according to any one of claims 1 to 16, the partition wall is made of a material that is permeable to gas but not permeable to liquid. And
[0030]
Further, the liquid sending mechanism according to claim 18 of the present invention is characterized in that, in the liquid sending mechanism according to claim 17, the material is a porous membrane having hydrophobicity.
Further, a liquid feeding mechanism according to a nineteenth aspect of the present invention is the liquid feeding mechanism according to the eighteenth aspect, wherein the porous film is formed of an organic polymer.
Furthermore, the liquid sending mechanism according to claim 20 of the present invention is characterized in that, in the liquid sending mechanism according to any one of claims 17 to 19, the outer surface of the partition wall is coated with a film that does not transmit gas and liquid. I do.
[0031]
Further, the liquid sending mechanism according to claim 21 of the present invention is characterized in that, in the liquid sending mechanism according to claim 20, the coating is bonded to the partition wall by thermocompression bonding or heat fusion.
Further, the liquid sending mechanism according to claim 22 of the present invention sends the liquid filled in a liquid tank formed by being surrounded by a wall to a flow path connected to the liquid tank, or Or a liquid sending mechanism for sending a liquid contained in another liquid tank connected to the flow path to the liquid tank, wherein a substance whose volume changes with a phase change is mixed with the liquid to be sent. It is characterized in that it is sealed inside the liquid tank so as not to prevent it.
[0032]
Further, the liquid sending mechanism according to claim 23 of the present invention is characterized in that, in the liquid sending mechanism according to claim 22, the substance is sealed inside the liquid tank while being separated from the liquid to be sent. And
Further, a liquid sending mechanism according to claim 24 of the present invention is characterized in that, in the liquid sending mechanism according to claim 22, the substance has a property of not mixing with the liquid to be sent.
[0033]
Further, in the liquid sending mechanism according to claim 25 of the present invention, in the liquid sending mechanism according to any one of claims 22 to 24, the temperature of the substance is controlled to generate a volume change due to a phase change in the substance. And at least one of a heating device and a cooling device.
With a liquid sending mechanism having such a configuration, accurate liquid sending can be performed even with a small amount of liquid. Further, the mechanism is simple, small, and low-cost, and can be suitably applied to an analyzer that performs various analyzes such as POC analysis.
[0034]
Further, if the partition wall is made of a material that is permeable to gas but not liquid, it acts as an air vent when filling the liquid in the liquid tank or the flow path, and deforms when the liquid is sent. Since the liquid can be pushed or pulled, a liquid feeding mechanism can be easily configured without using a special valve or mechanism.
In the present invention, the “outer surface of the partition” means a surface facing the outside of the liquid tank among the two surfaces of the partition.
[0035]
Furthermore, the analyzer according to claim 26 of the present invention is configured such that a liquid sample or a liquid mixture of a liquid sample and a liquid reagent is caused to flow into a flow path, and a predetermined amount of the sample or the liquid mixture is measured. An analyzer for analyzing a component, a chip having a liquid tank filled with the sample or the mixture, and the flow path connected to the liquid tank, and a chip according to any one of claims 1 to 25. And a liquid feeding mechanism described above.
[0036]
Further, in the analyzer according to claim 27 of the present invention, in the analyzer according to claim 26, the chip is formed by bonding a pair of flat members, and at least one of the pair of flat members is provided. One is provided with a groove on the plate surface, and the flow path is formed by bonding together with the plate surface having the groove inside.
[0037]
Furthermore, the analyzer according to claim 28 of the present invention is the analyzer according to claim 26 or claim 27, wherein the chip includes a liquid tank containing a dried reagent, and the liquid tank is provided in the liquid tank. The liquid reagent can be prepared in the chip by loading a lysis solution and dissolving the reagent with the lysis solution.
[0038]
With the analyzer having such a configuration, various analyzes including the POC analysis and the like can be performed easily and accurately in a short time. Further, since there is no need to provide a special device or mechanism for preparing a reagent solution necessary for analysis outside the chip, the configuration of the analyzer can be simplified.
Furthermore, the analyzer according to claim 29 of the present invention is the analyzer according to any one of claims 26 to 28, wherein the liquid in the plurality of liquid tanks or the plurality of channels is flushed with the liquid for liquid pushing. An analyzer provided with a mechanism for simultaneously moving the plurality of liquid tanks, wherein the plurality of liquid tanks include a liquid pushing channel through which the liquid for pushing pushed by the liquid sending mechanism according to any one of claims 1 to 25 flows. Alternatively, each of the plurality of flow paths is connected.
[0039]
With such a configuration, operations such as mixing and dilution of liquids necessary for analysis can be freely performed in the chip.
Hereinafter, the liquid feeding mechanism and the analyzer according to the present invention will be described in detail with reference to the drawings.
The liquid feeding mechanism of the present invention is mainly composed of two parts. That is, a portion having a liquid tank provided with a partition and a flow path (a portion corresponding to a chip, which is referred to as a chip in the following description), and a portion changing the volume of the liquid tank by deforming the partition (hereinafter, referred to as a chip). In the description, it is referred to as a liquid sending device). In order to realize a quantitative liquid transfer required for POC analysis or the like, it is necessary for the liquid transfer device to change the shape of the partition so that a volume change equal to a target liquid transfer amount is caused in the liquid tank. Therefore, it is necessary to accurately send a target amount of liquid according to a change in the volume of the liquid tank.
[0040]
[About chip]
The chip provided in the analyzer of the present invention includes a liquid tank filled with a liquid and a flow path through which a small amount of liquid is sent or reacted. The flow path is preferably formed by laminating a pair of flat members having grooves on the plate surface. That is, as shown in FIG. 1, a cover sheet 11 made of, for example, a resin is attached to a plate surface 10b provided with the groove 10a of a plate-shaped member 10 provided with a groove 10a on the plate surface via an adhesive, an adhesive tape or the like. When combined, a chip 1 having a capillary 12 serving as a liquid flow path is formed.
[0041]
This groove can be formed by a technique such as molding with a mold 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 path is about 0.3 to 10, and the width and the depth are Each is preferably 0.5 μm or more, and from the viewpoint of required sample and reagent amounts, the width and depth are each preferably 500 μm or less.
[0042]
When a resin is used as the material of the chip (plate-shaped member), it is required that the moldability is good, and when performing an optical measurement, the material must be transparent. It is preferred to use. Although glass can be used as the material of the chip, resin is preferable in consideration of cost.
Specifically, polystyrene, styrene-based resin such as styrene-acrylonitrile copolymer, methacrylic resin such as polymethyl methacrylate, methyl methacrylate-styrene copolymer, polycarbonate, polysulfone, polyether sulfone, polyetherimide, polyarylate, Examples include polymethylpentene and 1,3-cyclohexadiene-based polymers. It is also possible to use these copolymers and blends.
[0043]
[About the partition and liquid transfer using the partition]
As shown in FIG. 2A, the liquid tank 14 is formed by a through hole provided in the flat member 10, and an elastic partition wall 15 is attached so as to cover the opening. Since the partition wall 15 has flexibility and can be deformed, when a pressing force is applied to the partition wall 15 from the outside of the chip 1 as shown in FIG. Is pushed inside.
[0044]
Then, since the volume of the liquid tank 14 is reduced by the deformation of the partition wall 15, the liquid is incompressible. Therefore, as shown by an arrow in FIG. The capillary 12 is pushed out by the volume change. Thereby, the liquid L in the liquid tank 14 can be moved.
Thus, by performing the liquid sending using the deformation of the partition wall, desired liquid sending can be performed in the chip without using a complicated mechanism or equipment for connecting a pipe from the outside to the chip. In addition, since it is not necessary to provide dedicated equipment such as a pipe connection port on the chip, the chip can be manufactured easily and at low cost.
[0045]
The partition wall 15 is not particularly limited as long as it is a sheet-like member having flexibility and deformable by a small mechanism, but a PTFE (polytetrafluoroethylene) porous film is particularly preferable. If the partition wall 15 is formed of a film such as a PTFE porous membrane having a property of transmitting air but not liquid (repelled by water repellency), the partition wall 15 has gas permeability and the liquid tank 14 is formed of a liquid. When filled, it becomes a water-resistant functional film. 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 the liquid L is filled, and passes through the partition 15 to escape. Since the liquid L does not penetrate and escape due to the water repellency, the liquid tank 14 can be completely filled with the liquid L.
[0046]
In addition to PTFE, porous films of various materials can be used as the partition wall 15. However, it is preferable to use a hydrophobic organic polymer or 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., for example, polytetrafluoroethylene (PTFE), silicone, polyethylene, polypropylene, polystyrene, polyvinyl chloride. , Polycarbonate, polysulfone, polyethersulfone, polyarylate, polymethylpentene, 1,3-cyclohexadiene-based polymer and the like.
[0047]
Cellulose acetate membranes can be used in some cases, but in the case of a reagent solution to which a surfactant has been added, a membrane with high hydrophobicity, such as PTFE, silicone, or polyethylene, is more resistant to water permeation. Is preferred because it is large.
Since the liquid pressure can be sent at a higher pressure as the water pressure is higher, the water pressure is more preferable as the water pressure is higher. 0.01 MPa or more is preferable. However, when it is necessary to quickly fill the flow path or the liquid tank in the chip with liquid, the water pressure resistance is more preferably 0.1 MPa or more. The average pore diameter of the membrane can be from 0.1 μm to about 5 μm. However, considering that the smaller the pore diameter, the higher the water pressure resistance and the smaller the amount of permeated air, it is most preferably about 0.1 μm. The thickness is preferably 25 to 300 μm.
[0048]
As shown in FIG. 3, when the outer surface of the partition wall 15 is coated with a coating 16 that is impermeable to both gas and liquid, the water pressure resistance can be further increased. In the case where only the partition walls (PTFE porous membrane) do not have a coating, the gas in the liquid tank passes through the partition walls in the film thickness direction, whereas if the coating has the coating 16 as shown in FIG. The gas in the liquid tank passes through the inside of the membrane of the partition 15 toward the outer edge. Therefore, since the thickness of the partition wall 15 is the same as that of the partition wall 15, the water pressure resistance can be increased. When the partition walls are coated with a coating as described above, a membrane having an average pore diameter larger than about 0.1 μm described above can be used as the partition walls.
[0049]
When a PTFE porous film or the like having a high surface energy is used as a partition, it is generally difficult to bond the coating to the partition with an adhesive. In this case, if a thermoplastic resin such as low-density polyethylene is selected as the material of the coating, the coating can be bonded to a PTFE porous film or the like by thermocompression bonding or thermal fusion.
[About the liquid sending device]
First, an example of a liquid feeding device used in the liquid feeding mechanism according to claim 1 of the present invention is shown in a conceptual diagram of FIG. This liquid feeding device includes a piston 41, a non-compressible medium 42, a closed container 43, and a diaphragm member 44, which form part of the pressure generating means. The incompressible medium 42 is sealed in a closed space surrounded by the closed container 43 and the diaphragm member 44.
[0050]
As shown in FIG. 4B, pressure is applied to the incompressible medium 42 by the piston 41. At this time, the volume of the portion of the piston 41 existing in the incompressible medium 42 increases, and the amount of the increase is set to be equal to the target liquid supply amount. Then, the pressure is transmitted to the diaphragm member 44 via the incompressible medium 42 filled in the closed container 43, and the diaphragm member 44 is deformed so as to protrude toward the outside of the closed container 43. If the rigidity of the closed container 43 is sufficiently large, the volume of the protruding portion is equal to the above-described target liquid supply amount.
[0051]
Such a liquid feeding device 5a is mounted on the above-described chip 5b as shown in FIG. At this time, the outer surface of the diaphragm member 44 of the liquid feeding device 5a is brought into close contact with the outer surface of the partition wall 15 of the chip 5b.
As shown in FIG. 5 (b), when the pressure generating means is operated to apply pressure to the incompressible medium 42 by the piston 41, the diaphragm member 44 and the partition wall 15 are in close contact with each other. Due to the deformation of 44, the partition wall 15 is deformed so as to protrude toward the inside of the liquid tank 14. Then, since the volume of the liquid tank 14 is reduced by the deformation of the partition wall 15, the liquid L in the liquid tank 14 is sent out through the flow path 12 to the outside by the reduced volume.
[0052]
At this time, since the volume of the portion protruding into the liquid tank 14 is equal to the volume of the portion where the diaphragm member 44 protrudes, the liquid L having a volume equal to the target liquid supply amount described above is supplied. It becomes. That is, the pressure generated by the pressure generating means is transmitted to the partition 15 via the incompressible medium 42 and the diaphragm member 44, and the liquid L in the liquid tank 14 is sent to the outside by deforming the partition 15. . If the pressure generating means is operated so that a negative pressure is applied to the incompressible medium 42, the volume of the liquid tank 14 increases, so that the inside of the liquid tank 14 is It is also possible to draw the liquid L into the liquid. Thereby, the liquid in the flow path 12 or another liquid tank connected to the flow path 12 can be moved.
[0053]
Although not shown in FIGS. 4 and 5, the pressure generating means includes a drive source, and a linear actuator such as a linear step motor can be used as the drive source.
Examples of the incompressible medium 42 include liquids such as water and oil. In practice, the non-compressible medium 42 has properties such as silicone oil, which is non-volatile, liquid at room temperature, and chemically stable. It is desirable to have one. Instead of a liquid, a gel-like substance having sufficiently high fluidity can be used as the incompressible medium. In order to achieve the intended function in the present invention, it is necessary to prevent air bubbles from being mixed in the incompressible medium.
[0054]
Further, the diaphragm member 44 is made of a thin plate having elasticity and capable of holding the incompressible medium 42, and is preferably made of metal, resin, or the like. In order for the volume of the portion where the diaphragm member 44 protrudes and the change in the volume of the liquid tank 14 to be exactly equal, the incompressible medium 42 and the liquid in the liquid tank 14 which face each other via the diaphragm member 44 and the partition 15 are It is preferable that the shapes of the opposing surfaces are made equal and matched without deviation. However, when the outer edge of the diaphragm member 44 is supported as a fixed end, the deformation of the outer edge of the diaphragm member 44 is very small, so that the surface of the incompressible medium 42 and the liquid in the liquid tank 14 facing each other is small. Some deviation is allowed.
[0055]
In the liquid feeding mechanism shown in FIG. 5, it is assumed that the liquid feeding device 5a is used repeatedly and the chip 5b is replaced and used, but it is also possible to integrate both of them. In this case, as shown in FIG. 6, it is possible to configure a liquid feeding mechanism without using a diaphragm member.
Further, as shown in FIG. 7, the closed container 43 and the piston 41 which is a part of the pressure generating means may be connected by a pipe 70 (tubular member) filled with the incompressible medium 42. With such a configuration, it is possible to increase the degree of freedom in the layout design of the liquid sending mechanism and the analysis device using the liquid sending mechanism.
[0056]
4 to 7 adopt a reciprocating positive displacement pump mechanism (a positive displacement pump mechanism having a piston-cylinder mechanism or a plunger mechanism) as a pressure generating means, but another mechanism is used as a pressure generating means. May be adopted. For example, instead of the linear actuator and the reciprocating positive displacement pump mechanism, the pressure generating means may be configured by a rotary positive displacement pump mechanism such as a rotary motor and a gear pump.
[0057]
As a pressure generating means, that is, a means for causing a volume change, it is conceivable to use a volume change accompanying a phase change of a substance instead of the above-described one. For example, paraffin melts or solidifies at a temperature change from room temperature to several tens of degrees Celsius, at which time the volume increases or decreases by about 15%. If a small heater and a temperature sensor are arranged near the paraffin and the temperature of the paraffin is controlled, the volume change can be controlled.
[0058]
Volume change due to phase change includes volume change due to evaporation, and the degree of volume change is much larger than that in the case of solid-liquid phase change.However, since gas is highly compressible, it is necessary to send a fixed amount of liquid. In this case, it is not preferable.
FIG. 8A shows a liquid sending device using a volume change accompanying a phase change. In the closed container 43, the incompressible medium 42 and a substance 81 which changes in volume with a phase change are stored. The substance 81 which undergoes a volume change due to a phase change should be sealed in a bag or the like (not shown) which can expand and contract according to the volume change so as not to mix with the incompressible medium 42 when liquefied. Further, a heater 82 for heating is arranged in the bag. If a thermistor whose electric resistance changes according to the temperature is adopted as the heater 82 and the voltage drop at that time is measured by energizing the thermistor, a small and simple heating device having both a heater function and a temperature sensor function can be configured. It is possible to do.
[0059]
The volume change due to the phase change is transmitted to the diaphragm member 44 via the incompressible medium 42 without excess and deficiency, and the diaphragm member 44 protrudes by the amount of the volume change, and can function as the liquid sending device in the present invention. Note that if the substance 81 causing a volume change due to a phase change has a property of not mixing with the incompressible medium 42, the substance 81 does not need to be enclosed in a bag or the like. Further, the same function can be realized by filling the inside of the closed container 43 with a substance 81 that changes in volume with a phase change instead of the incompressible medium 42.
[0060]
The pressure generating means that focuses on the phase change of the substance does not have any mechanical element, and therefore can be easily miniaturized. Therefore, as shown in FIG. 8B, it is also possible to install the substance 81 which changes in volume with the phase change in the liquid tank 14 in the chip. As a heating means for causing a phase change, a method in which a substance 81 which causes a phase change together with the liquid tank 14 is heated by a heater 83 provided near the chip can be considered. Further, a method of heating a substance causing a phase change by laser condensing heating is also possible.
[0061]
The change in volume that determines the amount of liquid transfer appears as a change in the shape of the diaphragm member via the incompressible medium, and the change in volume is transmitted to the liquid in the liquid tank via the partition. The following requirements are required for accurate transmission of this volume change.
First, in order for the shape change of the diaphragm member to be accurately transmitted to the partition, it is desirable that the partition be as flexible as possible. Further, in order to exclude gas in the liquid tank when filling the liquid tank with the liquid, the partition wall needs to have a vent function. For example, a PTFE microporous membrane has this property.
[0062]
Second, it is preferable that there is no gap between the diaphragm member and the partition. For that purpose, first, the liquid sending device and the chip are mechanically overlapped. At this time, since the chip has a slight warp in manufacturing, the chip is mechanically pressed to enhance the close contact between them. However, since the partition wall is flexible and usually has a slack, there is a gap between the diaphragm member and the partition wall when viewed microscopically, which may not be negligible in precise quantitative liquid sending. In that case, it is necessary to make the outer surface of the partition wall and the outer surface of the diaphragm member adhere closely. Further, in order to facilitate the exchange of the chip, it is required that the state can be easily switched between the close contact state and the detached state.
[0063]
As a method of realizing switching between the close contact state and the detached state, there is a method using a vacuum as shown in FIG. That is, the vicinity of the diaphragm member 44 and the partition wall 15 is sealed using a gasket 91 or the like, and the pressure in the vicinity is reduced to a negative pressure using a vacuum pump or the like (not shown). If the liquid L in the liquid tank 14 is released to the atmospheric pressure through the flow channel 12, the partition wall 15 is pressed against and adheres to the surface of the diaphragm member 44 due to the pressure difference between the inner and outer surfaces of the partition wall 15. When the vacuum pump is stopped and the portion near the diaphragm member 44 and the partition 15 is returned to the atmospheric pressure, the liquid sending device and the chip are easily separated.
[0064]
In the case where the liquid feeding device is manufactured at a very low cost, and is integrated with the chip and is disposable, the diaphragm member and the partition wall may be bonded to each other so that no gap is generated between them.
[About liquid sending when an incompressible medium is composed of a substance having rubber elasticity]
In the present invention, it is ideal to employ a kind of miniature hydraulic mechanism as described above as the pressure generating means of the liquid feeding device. However, in order to actually manufacture a miniature hydraulic mechanism, precise processing and assembly are required, and it is not always easy to reliably remove bubbles from hydraulic oil. Therefore, it is possible to more easily configure the pressure generating means, the incompressible medium, and the diaphragm member, which are the components of the liquid feeding device of the present invention, by using rubber, which is an elastic body that can be regarded as substantially incompressible. it can.
[0065]
An example is shown in FIG. A rubber container 102 is filled in a cylindrical container 101 having one end open without any gap. A minute hole 104 is provided at the end of the container 101 on the closed side, and is connected to the minute hole 105 provided in the rubber 102. The pin 103 is inserted from the hole 104 to the hole 105.
When idealized and considered, since rubber is incompressible, the end face 106 of the rubber 102 expands by the volume of the inserted pin 103. If the rubber 102 is mounted on the chip so that the end face 106 and the partition of the chip are in close contact with each other, the change in shape of the end face 106 is transmitted to the liquid in the liquid tank via the partition. Then, the liquid is sent out to the flow path connected to the liquid tank by the volume calculated from the diameter of the pin 103 and the inserted length. In order for such a liquid sending mechanism to function correctly, the container 101 must have sufficient rigidity, and the diameters of the two should be selected so that no gap is formed between the hole 105 provided in the rubber 102 and the pin 103. It is necessary to do things.
[0066]
The holes 105 and the pins 103 provided in the rubber 102 do not necessarily have to be circular in cross section. For example, a similar function can be realized by providing a crack in the rubber 102 instead of the minute hole 105 and inserting a thin metal piece instead of the pin 103.
Further, it will be described that the liquid feeding mechanism using rubber as described above is also a displacement reduction mechanism. First, the case of a liquid feeding mechanism using rubber will be described. When a pin having a diameter of 0.5 mm is used, the cross-sectional area of the pin is 0.196 mm. ² When the insertion length is 5 mm, the change in volume, that is, the amount of liquid sent is 1.0 μL. That is, the insertion length corresponding to the liquid supply amount of 1.0 μL = the actuator drive amount is 5.0 mm.
[0067]
Next, in a case where the partition is directly pressed by a pin or the like, the actuator drive amount is roughly estimated. Assuming that the diameter of the partition wall is 3 mm and the shape of the partition wall deformed by the pin is a conical shape, the insertion length of the pin corresponding to 1.0 μL of the liquid supply amount is 0.43 mm. Therefore, the driving amount of the actuator is about 12 times larger in the case of the liquid feeding mechanism using rubber than in the case where the partition is directly pushed by a pin or the like. To put it simply, there is no problem even if the accuracy of the actuator drive amount is lower by an order of magnitude, so that the cost of the drive mechanism can be reduced.
[0068]
BEST MODE FOR CARRYING OUT THE INVENTION
An embodiment of a liquid sending mechanism according to the present invention and an analyzer provided with the liquid sending mechanism will be described in detail with reference to the drawings. In each drawing, the same or corresponding parts are denoted by the same reference characters. Note that the present embodiment is an example of the present invention, and the present invention is not limited to the present embodiment.
[0069]
[About the analyzer]
FIG. 11 is a conceptual diagram illustrating an overall configuration of an analyzer including all devices and devices necessary for analyzing a sample, such as a chip, a liquid feeder, an optical detector, and a temperature controller. The functions of the analyzer will be described below in the order of operation.
After the chip is introduced into the analyzer by the chip loading mechanism, the chip is sandwiched between the liquid sending device and the temperature controller, and the chip is fixed at a predetermined position. While operating the temperature control device to heat the chip, a preparation operation for introducing a buffer solution into an empty flow path and a liquid tank in the chip is performed using the time until the chip reaches a predetermined temperature. The liquid feeding device is operated to perform a dilution operation and a reaction operation as described below. The degree of reaction according to the concentration of the substance to be measured in the sample is measured using an optical detection device (for example, by absorbance). As the optical detection device, a photothermal conversion type detector based on the so-called thermal lens phenomenon can also be used, which is suitable for measuring the absorbance in a fine channel. The measurement process is controlled by the general CPU. In addition, the progress status and results of the measurement are appropriately presented to the measurer by the man-machine interface.
[0070]
With the analyzer having such a configuration, the analysis using the chip can be performed almost automatically and promptly.
[About chip]
An example of a chip used for an analyzer for biochemical measurement will be described with reference to the schematic diagram of FIG. The analysis process using this chip will be described later in detail.
[0071]
Several grooves each having a width of about 100 to 300 μm, a depth of about 50 μm, and a length of about 20 to 250 mm were formed on one surface of a transparent acrylic plate having a thickness of 2 mm by embossing. Further, through holes having a diameter of 4 to 5 mm were formed at both ends of these grooves. Then, a transparent acrylic sheet having a thickness of 0.3 mm was adhered to the plate surface having the groove of the plate using an ultraviolet curable adhesive, thereby forming a flow path.
[0072]
Further, a 70 μm-thick PTFE microporous membrane (average pore diameter: 0.1 μm) is attached to a portion of the other plate surface of the plate where the through hole is formed to cover an opening of the through hole. Thus, a partition was formed. With a commercially available PTFE microporous membrane, the required water pressure as a partition cannot be obtained, and the buffer solution in the liquid tank may infiltrate the PTFE microporous membrane and the valve function may be lost. If the outer surface of the porous film (the surface facing the outside of the chip) is covered with a thermoplastic resin such as a polyethylene thin film, the water pressure resistance can be improved. Even when coated with a thermoplastic resin, the function of removing air in the liquid tank was not inferior to the conventional one. The thermoplastic resin film is preferably bonded to the PTFE microporous film by thermocompression bonding or heat fusion.
[0073]
[About the liquid sending device]
A liquid transfer device capable of obtaining a liquid transfer amount of about 0.8 μL in a full stroke was manufactured as follows. An acrylic cylinder (outside diameter: 13 mm, inside diameter: 5 mm, height: 15 mm) having both ends open was filled with silicone rubber (Sylpot 184, manufactured by Dow Corning Japan) to a height of 10 mm, polymerized and solidified. The silicone rubber was filled with a two-part epoxy adhesive, and after solidification, a hole having a diameter of 1.0 mm was drilled to reach the upper end of the silicone rubber.
[0074]
Further, a needle having a diameter of 1.0 mm was inserted into the hole and inserted into the inside of the silicone rubber, thereby forming a crack-shaped hole for inserting and removing a pin described later in the rubber. Then, a pin having a diameter of 0.9 mm was inserted into the hole, and a linear actuator of 25 μm in one step was connected to this pin so that the pin could be inserted and removed from the silicone rubber.
[0075]
First, a measurement system shown in FIG. 13 was prepared to confirm the relationship between the insertion length of the pin and the protruding volume of the rubber in the above-described liquid sending device. In the measurement system of FIG. 13, the liquid feeding device 100 is arranged with the side into which the pin 103 is inserted downward, and the glass capillary 111 is attached to the rubber end surface 106 facing upward. Water is injected from the lower end side of the glass capillary 111 while taking care not to leave air bubbles, and the water is filled up to the middle of the entire length of the capillary. If the vertical movement of the water surface 112 when the pin 103 is inserted into the rubber 102 is measured by moving up and down a fine movement stage with a microscope (not shown), the measurement result and the cross-sectional area of the capillary 111 are used to determine the rubber end surface 106. Volume change due to protrusion can be calculated.
[0076]
Next, as shown in FIG. 14, the above-mentioned liquid sending device was arranged on the chip. At that time, the above-mentioned partition wall 15 of the chip and the diaphragm member of the liquid sending device were fixed so as to overlap. A gasket 91 made of silicone rubber is arranged around the acrylic container 101 of the liquid sending device, and a small vacuum pump (not shown) (not shown) for negative pressure in the vicinity of the partition 15 is connected to the partition 15. did.
[0077]
In the measurement system of FIG. 14, the liquid sending device 100 is attached to one end of the flow path 12 without branching or merging, and the glass capillary 111 is attached to the other end. Water is injected from the upper end of the glass capillary 111, air is released by the air permeability of the partition wall 15, and the injection of water is stopped when the liquid tank 14 becomes full. Water is drained from the lower end side of the glass capillary 111, and the water surface 112 is adjusted so as to be in the middle of the entire length of the capillary.
[0078]
Next, the small vacuum pump is operated to bring the partition 15 into close contact with the open surface of the rubber 102 of the liquid sending device. Then, by measuring the vertical movement of the water surface 112 when the pin 103 is inserted into the rubber 102 in the same manner as in the above-described measurement system, the relationship between the insertion length and the liquid sending amount is obtained.
The data thus obtained is shown in the graph of FIG. This graph shows the volume given by the product of the cross-sectional area of the pin 103 and the insertion length of the pin 103 as a theoretical value. As can be seen from the graph, the protruding volume of the rubber end face was a value several percent smaller than the theoretical value. This is considered to be because rubber is an approximate incompressible substance having a Poisson's ratio of about 0.49, and therefore has some compressibility. Is 0.5). The amount of liquid sent almost coincides with the protruding volume of the rubber end surface, indicating that the volume change is transmitted via the partition.
[0079]
When it is desired to change the relationship between the insertion length of the pin and the amount of liquid supplied, the cross-sectional area (diameter) of the pin may be changed. An example is shown in the graph of FIG.
[About liquid transfer mechanism using volume change due to phase change]
The pressure generating means can be constituted by using a substance which changes its volume during a phase change, and will be described below.
[0080]
Paraffin was used as the substance that caused a volume change during the phase change. As shown in FIG. 17, a container 151 having a volume of several tens of μL was connected to one end of a glass capillary 111. Then, 6 mg of paraffin 152 (manufactured by Wako Pure Chemical Industries, nominal melting point of 46 to 48 ° C.) was sealed in the container 151, and water was injected into the container 151 from the lower end side of the capillary 111. The amount of water to be injected was set so that the liquid level 112 reached the middle of the capillary 111, and care was taken to prevent bubbles from being mixed into the water as much as possible during water injection.
[0081]
The density of paraffin is about 1 g / cm ³ If the rate of volume change due to phase change is about 15%, melting of 6 mg of paraffin should result in about 1 μL of volume change. This change in volume is observed as a movement of the liquid surface 112 in the capillary 111. As shown in FIG. 17, a water tank 153 arranged so as to surround the container 151 is filled with water of about 70 ° C., and the change in the liquid level 112 of the capillary 111 when the temperature is gradually lowered by natural cooling is measured, and the volume change is performed. I asked.
[0082]
Actually, not only the paraffin but also the water, the container 151 and the capillary 111 are slightly changed in volume depending on the temperature, so that an error is caused in the measured value. Therefore, as a control experiment, the liquid level change was also measured for a system without paraffin, and the obtained data was subtracted from the data of the system with paraffin to correct the error.
[0083]
FIG. 18 is a graph showing the data of the volume change of the paraffin thus obtained due to the solid-liquid phase change. It was confirmed that about 1 μL of volume change occurred during the temperature change of 70 to 40 ° C.
Note that the above-described liquid feeding mechanism is an example of the liquid feeding mechanism according to claim 22 of the present invention. From the experiments described above, a substance such as paraffin, which changes its volume due to a solid-liquid phase change, is sealed in a liquid tank that is continuous with the flow path in the chip, and the substance undergoes a phase change by means such as heat transfer. It was shown that the liquid could be sent by doing so.
[0084]
However, in the above example, since paraffin has a property of not mixing with water, it is sealed with water in a liquid tank without being isolated from water, which is a liquid to be sent, but according to the volume change. If it is sealed in a liquid tank by separating it from the liquid to be sent by putting it in a bag that can be expanded and contracted, etc., even if it is a substance that mixes with the liquid to be sent, It can be adopted as a substance whose volume changes.
[0085]
[Unit operation required for analysis]
By combining a chip having a flow path with which the liquid flows and the liquid sending device, it is possible to perform operations such as liquid dilution and mixing, which are unit operations necessary for analysis. FIG. 19 shows an analytical device that performs only dilution prepared experimentally, and describes the result of evaluating the accuracy of the dilution.
[0086]
FIG. 19A is a cross-sectional view illustrating the configuration of a chip and a liquid feeding device fixed thereon. This liquid feeding device is one in which three liquid feeding devices using the aforementioned rubber are integrated. FIG. 19B is a plan view of the chip. Four liquid tanks are formed in this chip, and the P tank, the Q tank, and the Q 'tank are formed at the same pitch as that of the above-described triple liquid sending device.
[0087]
When the integrated pin 171 is pushed down and inserted into the hole formed in the rubber 172, the liquid L in the P tank, the Q tank, and the Q ′ tank is sent out to the flow paths connected to each tank, and the three flow paths are formed. At the junction.
A xylene cyanol dye simulating a sample to be measured is provided in the P tank, a flow path from the P tank to the junction, and a flow path downstream of the junction (post-merge flow path). An aqueous solution (concentration: 400 μM) is filled. Further, the Q tank, the Q ′ tank, the flow path from the Q tank to the junction, and the flow path from the Q ′ tank to the junction are filled with a buffer solution.
[0088]
When the liquid sending device is operated, the aqueous dye solution and the buffer solution merge. Then, a dilution operation is performed by the two being diffused and mixed in the flow path. Here, the diameter of the buffer solution pin is 0.7 mm, and the diameter of the sample sample (xylene cyanol dye aqueous solution) pin is 0.3 mm. However, if the dilution rate needs to be changed, it is necessary to correspond to each liquid tank. What is necessary is just to change the diameter of a pin.
[0089]
If a photothermal conversion type absorbance measurement system (thermal lens measurement system) is arranged near the end of the flow path after the merging, the change in the concentration of the liquid passing through the end can be detected, so that the dilution ratio can be measured. is there.
FIG. 20 shows the result of measuring the dilution ratio. This graph shows how the absorbance of the liquid decreases due to dilution after the start of liquid supply. The results of numerical calculations by differentially approximating the advection-diffusion phenomenon based on separately measured characteristics of the liquid sending device are shown as calculated values.
[0090]
As can be seen from the graph, the experimental value and the calculated value were almost the same, and it was confirmed from this that the intended dilution operation could be realized with this liquid sending mechanism.
[About analysis operation]
Next, the analysis process using the chip will be described in detail. FIG. 12 is a schematic diagram illustrating the configuration of a chip of an analyzer for biochemical measurement. This chip is provided with 9 liquid tanks (P tank, Q tank, Q 'tank, D tank, S tank, A tank, B tank, R tank, W tank), Are in communication.
[0091]
The P tank is filled with plasma separated from blood, and the A and B tanks have dried reaction reagents attached to the bottom of the tank. The R tank is filled with a buffer solution. Further, the opening of each liquid tank is covered with a partition wall (a porous PTFE membrane having a pore size of about 0.1 μm and a water pressure resistance of 0.39 MPa) having flexibility, gas permeability, and water resistance. The P tank, the Q tank, and the Q ′ tank are equipped with a triple liquid feeding device shown in FIG. Further, a liquid feeding device 100 as shown in FIG. 14 is mounted in each of the D tank and the R tank. Further, the valve 201 shown in FIG. 21 is attached to the W tank, and the valve 201 is closed in an initial state. The valve 201 is installed so as to seal the periphery of the partition 15 of the W tank with a gasket 202. When the valve 201 is open, the air in the W tank can pass through the partition and flow out of the valve 201, and when the valve 201 is closed, the air in the W tank passes through the partition and the valve 201 It is prevented from flowing out of the. Therefore, by opening or closing the valve 201, it is possible to select whether or not to replace the air in the W tank with the liquid.
[0092]
First, the liquid sending device attached to the R tank is operated to fill the entire flow path in the chip with the buffer solution. That is, the buffer solution filled in the R tank is pushed out by the liquid sending device arranged in the R tank, but since the valve 201 of the W tank is in the closed state, the buffer solution flows through the flow paths sab, sa, and sab. s flows into the S tank, the A tank, and the B tank.
When the S tank is full, the buffer flows into the Q tank and Q 'tank, but does not flow into the P tank because the P tank is already filled with the sample. When the tank A and the tank B are full, the buffer flows into the tank D. Finally, the filling of the buffer solution into the Q tank, Q ′ tank, D tank, S tank, A tank, and B tank is completed. Since the valve 201 is closed in the W tank, only the buffer solution passes therethrough, and the inside of the W tank is almost filled with air.
[0093]
In this process, the dried A reagent and the dried B reagent stored in the A and B tanks are dissolved in the buffer solution, and thus the respective reagent solutions are prepared in the A and B tanks. The concentration of the reagent solution can be set to a desired value by the volume of the liquid tank and the amount of the dry reagent. By such an operation, the dissolution of the reagent and the filling of each liquid tank with the buffer solution for sending the liquid were completed.
[0094]
Next, the sample is diluted. When the valve 201 of the W tank is opened and the sample and the buffer solution are simultaneously pushed out by the liquid sending devices mounted on the P tank, the Q tank, and the Q ′ tank, the two liquids diffuse and mix, and the mixed liquid is mixed. It flows into the S tank. That is, the buffer in the S tank is replaced with the diluted sample. The buffer initially present in the S tank flows into the W tank, which is a waste liquid tank, through the flow path s, the flow path sa, and the flow path sab. Note that the dilution ratio of the sample is determined by the set flow rate of the liquid sending device mounted on the P tank, the Q tank, and the Q ′ tank.
[0095]
At this stage, the tank S was filled with the diluted sample, and the tanks A and B were filled with the respective reagent solutions, and the preparation for the reaction was completed.
Finally, when the buffer solution filled in the D tank is pushed out by the liquid sending mechanism mounted on the D tank, the buffer solution flows into the S tank, the A tank, and the B tank via the three branch flow paths. The liquid content of each liquid tank is sent out to each flow path connected to the liquid tank. That is, the diluted sample is sent to the channel s, the reagent A is sent to the channel a, and the reagent B is sent to the channel b. At this time, the buffer acts as a liquid for pushing.
[0096]
The flow rates are distributed to the respective flow paths so that the pressure loss of each flow path at the branch point of the three branch flow paths becomes equal. In designing the chip, the pressure loss coefficient of each flow path is determined in consideration of the difference in the viscosity of each liquid so that the flow rates of the diluted sample and each of the reagent solutions A and B become desired values. The pressure loss coefficient is determined by two factors: the shape of the cross section of the flow channel and the length of the flow channel. Since the liquid feeders mounted in the P, Q and Q 'tanks are stopped, no liquid flows into these liquid tanks.
[0097]
The diluted sample and the A reagent solution flow into the flow channel sa and are mixed, and a reaction between the sample and the A reagent occurs in the flow channel sa. The mixed solution flows toward the junction with the flow channel b while reacting, and the reaction time can be controlled by the flow speed and the length of the flow channel sa, when the time flowing through the flow channel sa is defined as the reaction time.
After the reaction between the sample and the reagent A has progressed for a predetermined time, the mixture is made to reach the mixing point with the flow channel b. Thereby, the mixed solution with which the final concentration measurement can be performed flows into the flow channel sab by further mixing with the reagent B and causing a reaction. After mixing, after an appropriate reaction time compatible with the reagent reaction system, labeling is performed as a dye concentration. If the length of the flow channel sab is set so that a sufficient reaction time can be taken in consideration of the flow rate, immediately after the reaction, the optical measurement device as described in FIG. Can be measured.
[0098]
The mixed solution whose concentration has been measured reaches a W tank (waste liquid tank) and is stored as a waste liquid. Since the chip after the concentration measurement can be disposable as it is, it is possible to construct an analysis system that does not cause a specimen-derived infection or the like to an analysis operator.
In addition, the dilution and extraction processes of a sample such as blood can be considered in the same manner, and if the above-described method is used, almost all possible reagent reaction processes can be realized.
[0099]
【The invention's effect】
As described above, the analyzer of the present invention can easily and accurately perform various analyzes including POC analysis. Further, the liquid sending mechanism of the present invention is suitable for sending liquid in the analyzer, has a small and simple configuration, and is low in cost.
[Brief description of the drawings]
FIG. 1 is a cross-sectional view illustrating a configuration of a chip of an analyzer according to the present invention.
FIG. 2 is a schematic diagram illustrating the principle of liquid feeding in the present invention.
FIG. 3 is a schematic diagram illustrating a means for improving the water resistance of a partition.
FIG. 4 is a schematic diagram illustrating the principle of a liquid sending device.
FIG. 5 is a schematic diagram illustrating the principle of performing liquid sending by combining a liquid sending device and a chip.
FIG. 6 is a schematic diagram illustrating another liquid feeding mechanism according to the present invention.
FIG. 7 is a schematic diagram of a liquid sending device having a configuration in which a closed vessel and pressure generating means are connected by a pipe.
FIG. 8 is a schematic diagram illustrating a liquid sending mechanism using a substance that changes in volume with a change in solid-liquid phase as pressure generating means.
FIG. 9 is a schematic diagram illustrating a means for securely bringing a diaphragm member and a partition into close contact with each other.
FIG. 10 is a schematic diagram of a liquid sending device in which an incompressible medium is made of rubber.
FIG. 11 is a schematic diagram showing the overall configuration of an analyzer according to one embodiment of the present invention.
FIG. 12 is a schematic diagram illustrating a channel configuration of a chip for performing a sample diluting operation and a reaction operation.
FIG. 13 is a schematic diagram illustrating a method of measuring a liquid sending amount by a liquid sending device.
FIG. 14 is a schematic diagram illustrating a method for measuring a liquid sending amount by a liquid sending device and a chip.
FIG. 15 is a graph showing a result of measuring a correlation between a pin insertion length and a liquid sending amount in the liquid sending device and the chip of FIG. 14;
FIG. 16 is a graph showing the results of measuring the correlation between the pin insertion length and the amount of liquid supplied while changing the diameter of the pin.
FIG. 17 is a schematic diagram illustrating a method of measuring a liquid sending amount in a liquid sending mechanism using a substance that changes in volume with a solid-liquid phase change as pressure generating means.
18 is a graph showing the results of measuring the characteristics of the liquid sending mechanism of the liquid sending mechanism of FIG.
FIG. 19 is a schematic diagram illustrating a dilution operation performed using a liquid sending device and a chip.
FIG. 20 is a graph showing the results of measuring characteristics of a dilution operation performed using the liquid sending device and the chip of FIG. 19;
FIG. 21 is a schematic diagram showing a structure of a valve used in a preparatory operation when performing a sample diluting operation and a reaction operation.
[Explanation of symbols]
1,5b chip
10 flat members
10a groove
10b Plate surface
11 Cover sheet
12 Capillary (flow path)
14 Liquid tank
15 diaphragm
16 Coating
41 piston
42 Incompressible medium
43 sealed container
44 Diaphragm member
70 pipe
81 Substance that causes volume change with phase change
82, 83 heater
101 container
102,172 rubber
103,171 pins
104 holes
105 holes
152 paraffin
L liquid

Claims (29)

By changing the volume of the liquid tank formed by being surrounded by the wall, the liquid filled in the liquid tank is sent to a flow path connected to the liquid tank, or the flow path or the flow path A liquid sending mechanism for sending a liquid stored in another connected liquid tank to the liquid tank,
At least a part of the wall body is formed of a partition having elasticity that can be deformed so as to project toward the inside or outside of the liquid tank,
A diaphragm member in contact with the outer surface of the partition wall, a non-compressible medium in contact with a surface of the diaphragm member that is not in contact with the partition wall, a container storing the non-compressible medium, and the incompressible medium. Pressure generating means for applying pressure, the incompressible medium is sealed in a closed space surrounded by the diaphragm member and the container,
The liquid feeding mechanism, wherein the pressure is applied to the partition via the incompressible medium and the diaphragm member, so that the partition is deformed. The liquid feeding mechanism according to claim 1, wherein the container and the pressure generating unit are connected by a tubular member filled with an incompressible medium. The liquid sending mechanism according to claim 1, wherein the pressure generating unit generates the pressure using a reciprocating positive displacement pump mechanism. The liquid sending mechanism according to claim 1, wherein the pressure generating unit generates the pressure using a rotary positive displacement pump mechanism. The liquid sending mechanism according to claim 1, wherein the pressure generating unit generates the pressure using a volume change accompanying a phase change of a substance. The liquid sending mechanism according to claim 5, further comprising at least one of a heating device and a cooling device that controls a temperature of the substance to cause a volume change of the substance due to a phase change. The incompressible medium is made of a material having rubber elasticity, and the pressure generating means is formed of the container provided with a hole or a crack, and the non-compressible medium provided with a hole or a crack continuous with the hole or the crack of the container. 2. The liquid feeding mechanism according to claim 1, comprising a compressible medium, and a pin or a flake inserted in holes or cracks of the container and the incompressible medium and reciprocated. 3. By reducing the pressure near the contact portion between the partition and the diaphragm member to less than the atmospheric pressure, the partition and the diaphragm member are brought into close contact with each other, and the pressure near the contact portion is set to the atmospheric pressure, whereby the partition and the diaphragm member are reduced. The liquid feeding mechanism according to any one of claims 1 to 7, further comprising: means for releasing contact with the liquid supply mechanism. By changing the volume of the liquid tank formed by being surrounded by the wall, the liquid filled in the liquid tank is sent to a flow path connected to the liquid tank, or the flow path or the flow path A liquid sending mechanism for sending a liquid stored in another connected liquid tank to the liquid tank,
At least a part of the wall body is formed of a partition having elasticity that can be deformed so as to project toward the inside or outside of the liquid tank,
An incompressible medium that is in contact with the outer surface of the partition, a container that stores the incompressible medium, and a pressure generator that applies pressure to the incompressible medium, wherein the incompressible medium includes the partition and Sealed in a closed space surrounded by the container,
The liquid feeding mechanism, wherein the pressure is applied to the partition via the incompressible medium, and the partition is deformed. The liquid sending mechanism according to claim 9, wherein the container and the pressure generating means are connected by a tubular member filled with an incompressible medium. The liquid sending mechanism according to claim 9, wherein the pressure generating unit generates the pressure using a reciprocating positive displacement pump mechanism. The liquid sending mechanism according to claim 9, wherein the pressure generating unit generates the pressure using a rotary positive displacement pump mechanism. The liquid sending mechanism according to claim 9, wherein the pressure generating unit generates the pressure using a volume change accompanying a phase change of a substance. 14. The liquid feeding mechanism according to claim 13, further comprising at least one of a heating device and a cooling device for controlling a temperature of the substance to cause a volume change of the substance due to a phase change. The incompressible medium is made of a material having rubber elasticity, and the pressure generating means is formed of the container provided with a hole or a crack, and the non-compressible medium provided with a hole or a crack continuous with the hole or the crack of the container. The liquid feeding mechanism according to claim 9, comprising a compressible medium, and pins or flakes that are inserted into holes or cracks of the container and the incompressible medium and reciprocate. The liquid sending mechanism according to any one of claims 9 to 15, wherein at least a part of a contact portion between the partition and the incompressible medium is adhered. The liquid feeding mechanism according to any one of claims 1 to 16, wherein the partition wall is made of a material that is permeable to gas and not permeable to liquid. The liquid feeding mechanism according to claim 17, wherein the material is a porous membrane having hydrophobicity. The liquid feeding mechanism according to claim 18, wherein the porous film is formed of an organic polymer. 20. The liquid feeding mechanism according to claim 17, wherein an outer surface of the partition wall is coated with a film that does not allow gas and liquid to pass through. 21. The liquid feeding mechanism according to claim 20, wherein the coating is bonded to the partition wall by thermocompression bonding or thermal fusion. The liquid filled in the liquid tank formed by being surrounded by the wall is sent to a flow path connected to the liquid tank, or is stored in the flow path or another liquid tank connected to the flow path. A liquid sending mechanism for sending the liquid to the liquid tank,
A liquid sending mechanism characterized in that a substance whose volume changes with a phase change is sealed inside the liquid tank so as not to mix with the liquid to be sent. 23. The liquid sending mechanism according to claim 22, wherein the substance is sealed inside the liquid tank while being separated from the liquid to be sent. 23. The liquid sending mechanism according to claim 22, wherein the substance has a property of not mixing with the liquid to be sent. The liquid feeding mechanism according to any one of claims 22 to 24, further comprising at least one of a heating device and a cooling device that control a temperature of the substance to cause a volume change of the substance due to a phase change. A liquid sample, or an analyzer for flowing a mixture of a liquid sample and a liquid reagent into a flow path, and analyzing a predetermined component in the sample or the mixture,
A liquid tank filled with the sample or the mixed liquid, and a chip having the flow path connected to the liquid tank,
A liquid feeding mechanism according to any one of claims 1 to 25,
An analyzer comprising: The chip is formed by laminating a pair of flat members, and at least one of the pair of flat members has a groove in a plate surface, and is bonded with the grooved plate surface inside. The analyzer according to claim 26, wherein the flow path is formed. The chip is provided with a liquid tank in which a dried reagent is stored, and a dissolving solution is charged into the liquid tank and the reagent is dissolved in the dissolving solution. 28. The analyzer according to claim 26, wherein the analyzer can be prepared by the following method. An analyzer having a mechanism for simultaneously moving a plurality of liquid tanks or liquids in a plurality of flow paths by flushing the liquid in the liquid for liquid pushing,
A liquid pushing flow path through which the liquid pushing liquid sent by the liquid sending mechanism according to any one of claims 1 to 25 is connected to the plurality of liquid tanks or the plurality of flow paths, respectively. An analyzer according to any one of claims 26 to 28.

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