JP2006023121A - Liquid feeder - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a small-sized inexpensive liquid feeder suitably used in various analyzers inclusive of a POC analyzer and the like. <P>SOLUTION: The liquid feeder is equipped with a liquid housing body having a liquid storage tank and a uncompressible medium housing body for housing an uncompressible medium for transmitting pressure for causing the volumetric change of the liquid storage tank. A diaphragm deformable so as to protrude toward the inside or outside of the liquid storage tank is provided on the liquid storage tank. Further, the uncompressible medium housing body is equipped with a first medium storage tank equipped with a transforming means for producing pressure and filled with the uncompressible medium and a second medium storage tank allowed to communicate with the first medium storage tank by an uncompressible medium flow channel. The liquid storage tank and the second medium storage tank are arranged adjascent to each other so as to hold the diaphragm and the diaphragm is deformed so as to follow a change in the pressure applied to the uncompressible medium by the transforming means. <P>COPYRIGHT: (C)2006,JPO&amp;NCIPI

Description

  The present invention relates to a liquid feeding device that is suitably used for an analyzer that easily analyzes and detects a trace amount of sample.

  Analyzes for bedside diagnosis (POC (point of care) analysis) that performs measurements necessary for medical diagnosis in the vicinity of patients, and analyzes of hazardous substances in rivers and wastes at rivers and waste disposal sites (POU (point of use) analysis), and analysis and measurement at or near the site where analysis / measurement is required, such as contamination inspection at food cooking, harvesting and import sites The importance of (hereinafter collectively referred to as “POC analysis, etc.”) has attracted attention. In recent years, the development of detection methods and apparatuses applied to such POC analysis and the like is being emphasized. Such POC analysis and the like are required to be performed simply in a short time and at a low cost.

  Conventionally, for microanalysis, a GC-MS apparatus or an LC-MS apparatus, in which a sample is separated by capillary gas chromatography (CGC), capillary liquid chromatography (CLC), etc. and then quantified by a mass spectrometer, has been widely used. It was. However, these analyzers are large in mass spectrometers and cumbersome to operate, so they should not be used for on-site measurements such as patient bedsides, contaminated rivers, and near waste disposal sites. Not suitable. Furthermore, it is desirable that an analyzer for medical diagnosis using blood or the like as a sample should be disposable at the part touched by the sample.

  Therefore, in order to solve these problems, studies have been made to reduce the size of a conventionally used analyzer and apply μTAS (micro total analysis system) technology for reacting a very small 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 centimeters or less in order to reduce the amount of a specimen, not limited to blood, and a reagent solution or specimen is allowed to flow through the groove. A small amount of sample is analyzed by performing separation and reaction (see, for example, Patent Documents 1, 2, and 3 and Non-Patent Document 1). This technology has the advantage that the amount of specimen, 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 reduced, and the time required for detection can be reduced to a short time. is there.

The applicant of the present application also describes “mixing analysis apparatus and mixing analysis method” (Patent Document 4), “capillary photothermal conversion analysis apparatus” (Patent Document 5), “analysis apparatus” (Patent Document 6), “analysis cartridge and liquid feeding We have filed an application related to μTAS, such as “Control Device” (Patent Documents 7 and 8).
These publications or application specifications also describe using a resin microchip as a chip and using a thermal lens detection method as a trace component detection method.

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. (Patent Documents 9, 10, 11 and Non-Patent Documents 2, 3, 4, 5, 6, etc.).
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 high sensitivity can be realized without an operation such as introduction of a fluorescent labeling substance. Thermal lens detection is suitable.

On the other hand, there are also proposed methods for supplying liquid by applying voltage using capillary electrophoresis itself or electroosmotic flow in a chip (Patent Documents 12, 13, 14 and Non-Patent Documents 3, 7, etc.).
However, since both electrophoresis and electroosmotic flow apply a voltage to the liquid in the chip via the electrode, the reagent and sample composition change due to electrolysis of the measurement reagent and measurement sample on the electrode surface. There is. In addition, a phenomenon may occur in which the electrolytic product of the reagent or sample adheres to the inner surface of the capillary, changes the zeta potential on the capillary surface, and changes the liquid feeding speed.

In addition, a solid reagent freeze-dried is placed in the cartridge, the blood sample is diluted with a lysis dilution solution enclosed in the cartridge, and the solid reagent is further dissolved in the diluted sample solution, and an analytical reaction is performed for quantification. A method is disclosed (Patent Documents 15, 16, etc.).
In this method, since liquid feeding is performed by centrifugal force, the liquid feeding direction is always the direction in which the centrifugal force works, that is, the direction from the center of the circle of the circular cartridge to the outside. 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 specimen flows into each small chamber, dissolves the solid reagent, reacts, and changes to absorbance. Come to come.

  However, because of the structure of the cartridge, since the solid reagent is placed at the final point of the flow path, only a one-step detection reaction with one reagent can be performed. Depending on the detection item, a clinical laboratory in a test center or hospital Therefore, it is necessary to adopt a reaction and reagent composition different from the detection reaction based on the recommended method established by academic societies or government offices. Therefore, the correlation with conventional inspection data may be low. Further, depending on the inspection item, it may be difficult to analyze in such a circular cartridge reaction mode.

  On the other hand, a liquid feeding method using bubbles in a sealed capsule has been reported. This includes liquid feeding due to thermal expansion of bubbles and liquid feeding due to generation of electrolytic gas. The former is performed by a sealed chamber, a heating material, and a filling liquid. The expansion of the heated bubbles generates a high pressure and enables fast liquid feeding. For example, in the method of Naruse Yoshihiro et al., If a light-absorbing heat generating material is sealed in a chamber filled with liquid and the laser light is guided into the chamber with glass fiber, the bubbles in the chamber expand due to the heat generation of the light-absorbing heat generating material. Thus, the liquid is fed (Patent Document 17).

On the other hand, the latter is performed by a sealed chamber, an electrolytic solution filled therein, and a pair of electrodes inserted in the chamber. Pressure is generated by the generation of electrolytic gas by applying a voltage to the electrodes, and liquid feeding is performed. For example, there are those reported by DAHopkins, Jr (patent document 18) and CRNeagu et al. (Non-patent document 8).
However, since bubbles have compressibility, when a liquid is sent into a narrow capillary channel, it takes a long time to stabilize after the liquid flow starts. In some cases, the liquid may vibrate in the flow direction, which may cause a problem that the liquid feeding is not stabilized at all.

Furthermore, a method of sending the liquid by pushing the filling liquid from the outside without using bubbles has been proposed, for example, liquid feeding by a piezoelectric element. A piezoelectric element can generate a large force with a relatively small amount of energy.
However, when the piezoelectric element is composed of a single crystal, the liquid can be pushed and moved only by a very small distance. In order to increase the stroke, the piezoelectric element is usually constituted by a plurality of crystals. However, if this is done, a large number of parts are required, resulting in an increase in cost.

  In addition, since the piezoelectric element is driven with a small current but requires a high voltage, it is not necessarily adapted to the present semiconductor circuit. Furthermore, it is necessary to form piezoelectric elements by laminating materials having different expansion coefficients, and accurate clearance is required for lamination, so it is difficult to reduce the size. Furthermore, because it is a reciprocating motion due to vibration, in order to convert it into a unidirectional force suitable for liquid feeding, it is necessary to use multiple valves with check valve functions or to add phase differences to multiple pumps. There is a problem that the entire system becomes very complicated because control is required.

Although it is possible to perform liquid feeding by installing a module having a rectifying effect such as a diffuser in the flow path, the rectifying effect cannot be expected unless the flow rate is high due to its structural characteristics. Although it is possible to increase the speed by narrowing the flow path width at low flow rates, this method increases the pressure loss in the flow path, and it is necessary to improve the chip manufacturing accuracy and flow control accuracy. It becomes difficult to build a practical system for reasons such as high costs.
Furthermore, liquid feeding based on the principle of electrostatic charge repulsion and counter charge is reported. For example, in the method of R. Zengerle et al., The liquid in the chamber is pushed out by electrostatic repulsion between the thin membrane electrode and the chip fixing electrode (Non-patent Document 9).

However, since the gap distance is sensitive to 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 in a high electric field. Such contamination prevents proper liquid feeding. Moreover, since a large current is not required but a high voltage is required, it is not necessarily adapted to today's semiconductor circuits. Furthermore, a large capacity charged plate is required to obtain a large liquid feeding force. 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 feeding. There is a problem that the whole becomes very complicated.
JP-A-2-245655 JP-A-3-226666 JP-A-8-233778 Japanese Patent Application No. 10-181586 JP 2000-2675 A Japanese Patent Laid-Open No. 2000-2777 International Publication No. 99/64846 Pamphlet Japanese Patent Application No. 11-227624 JP 60-174933 A Japanese Patent Laid-Open No. 10-142177 JP-A-4-369467 International Publication No. 96/04547 Pamphlet Japanese Patent Laid-Open No. 10-142177 JP-A-4-369467 Japanese National Patent Publication No. 10-501340 JP 9-504732 A US Pat. No. 5,210,817 US Pat. No. 5,671,905 Analytical Chem. 69, 2626-2630 (1997) AC Boccara et. Al., Appl. Phys. Lett. 36, 130, 1980 J. Liquid Chromatography 12,2575-2585 (1989) M. Harada et. Al. 4,280-284,1997 Anal. Chem. Vol.65,2938-2940,1993 Kawanishi et al., Abstracts of the 44th Annual Meeting of the Analytical Society of Japan, p119, 1995 SC Jakobson et.al., Anal. Chem. Vol.66,4127-4132,1994 CRNeagu, JGEGardeniers, M. Elwenspoek, JJKelly, Journal of Micro Electro Mechanical Systems, 5,1, 2-9, (1996) R. Zengerle, A. Richter, H. Sandmaier, Micro Electro Mechanical Systems '92, 4, 19, (1992)

  Devices that perform POC analysis or the like (hereinafter may be referred to as POC analysis devices) generally have requirements such as small size, low cost, simple operation, and short measurement time. For example, a POC analyzer used in a small medical institution such as a clinic cannot be installed in a limited space in a clinic, has a small equipment size, a low cost of capital investment, and is not a specialized laboratory technician. However, it is necessary to satisfy the requirements such as the ease of operation and the ability to measure within several tens of minutes without waiting for the patient. It is also necessary to be able to measure multiple items simultaneously for accurate diagnosis of medical conditions, and to be able to handle quantitative measurements.

As mentioned above, many proposals have been made as liquid feeding technology to be provided to POC analyzers. However, all of these requirements for small size, low cost, simplicity, short measurement time, multi-item measurement, and quantitativeness are required. No match has been proposed yet.
Accordingly, an object of the present invention is to solve the above-described problems of the prior art, and to provide a liquid feeding device that can be easily manufactured without being complicated and easy to manufacture. It is to be.
Another object of the present invention is that the specimen or reagent stays only in the disposable cartridge, does not touch any part other than the cartridge of the liquid feeding device, does not generate contamination, and is affected by the properties of the specimen and reagent. It is an object of the present invention to provide a liquid delivery device that can deliver liquid quantitatively without any problems.

  In order to solve the above problems, the present invention has the following configuration. That is, in the liquid feeding device according to claim 1 of the present invention, the liquid filled in the liquid storage tank is connected to the liquid storage tank by changing the volume of the liquid storage tank formed surrounded by the wall. A liquid feeding device that sends the liquid stored in the flow path or another storage tank connected to the flow path to the liquid storage tank, the liquid container having the liquid storage tank; and An incompressible medium container that contains an incompressible medium that transmits a pressure for causing a volume change of the liquid storage tank, and the liquid storage tank includes at least a part of the wall body of the liquid storage tank. The incompressible medium container includes a transforming means that generates the pressure and is filled with the incompressible medium. One medium storage tank and incompressible A second medium storage tank connected to the first medium storage tank by a body flow path, and the liquid storage tank and the second medium storage tank are disposed adjacent to each other with the diaphragm interposed therebetween, The deformation means is adapted to deform following the pressure change applied to the incompressible medium.

  According to a second aspect of the present invention, in the liquid delivery device according to the present invention, the liquid filled in the liquid storage tank is connected to the liquid storage tank by changing the volume of the liquid storage tank formed by being surrounded by the wall. A liquid feeding device that sends the liquid stored in the flow path or another storage tank connected to the flow path to the liquid storage tank, the liquid container having the liquid storage tank; and An incompressible medium container that contains an incompressible medium that transmits a pressure for causing a volume change of the liquid storage tank, and the liquid storage tank includes at least a part of the wall body of the liquid storage tank. The incompressible medium container includes a transforming means that generates the pressure and is filled with the incompressible medium. One medium storage tank and incompressible medium A second medium storage tank connected to the first medium storage tank by a path, and the liquid storage tank and the second medium storage tank are provided in the second medium storage tank so as to be in contact with the diaphragm and the diaphragm. The diaphragm member is arranged adjacent to the diaphragm member, and the diaphragm member follows the pressure change applied to the incompressible medium by the transformation means and protrudes toward the inside or the outside of the liquid storage tank. The diaphragm can be deformed according to the deformation of the diaphragm member.

Furthermore, in the liquid feeding device according to claim 3 according to the present invention, in the liquid feeding device according to claim 1 or 2, the liquid container is configured by bonding a pair of flat plate members, At least one of the pair of flat plate-like members is provided with a groove on a plate surface, and the flow path is formed by bonding the plate surface provided with the groove inside.
Furthermore, the liquid feeding device according to a fourth aspect of the present invention is the liquid feeding device according to the first or second aspect, wherein the incompressible medium container is formed by bonding a pair of flat plate members. And at least one of the pair of flat plate-like members is provided with a groove on a plate surface, and the incompressible medium flow path is formed by bonding the plate surface provided with the groove inside. Features.

  With such a configuration, the pressure generated by the transformer means is transmitted to the diaphragm via the incompressible medium, and the diaphragm is deformed so as to protrude toward the inside or the outside of the liquid storage tank. Since the volume of the liquid storage tank changes due to the deformation of the diaphragm, the liquid filled in the liquid storage tank is sent to the flow path, or the liquid stored in the flow path or another storage tank is the liquid storage tank. The liquid is sent to The flow rate of the liquid at this time does not depend on the properties of the liquid and can be controlled by the deformation amount of the diaphragm (that is, the amount of transformation applied to the incompressible medium), so that the liquid in the liquid container is quantitatively sent. It is possible to liquid.

  Since the liquid feeding device of the present invention can control the flow rate of the liquid according to the deformation amount of the diaphragm, the liquid can be quantitatively fed.

DESCRIPTION OF EMBODIMENTS Hereinafter, embodiments of a liquid feeding device and an analyzer equipped with the liquid feeding device according to the present invention will be described with reference to the drawings. In the following drawings, the same or corresponding parts are denoted by the same reference numerals. Moreover, this embodiment shows an example of this invention, Comprising: This invention is not limited to this.
In FIG. 1, the top view of an incompressible medium container and AA 'sectional drawing of this top view are shown. The incompressible medium container 1 has a plate-like structure, and includes a first medium storage tank 8, second medium storage tanks 10, 11, and 12, and a first medium storage tank 8 and second medium storage tanks 10, 11, and 12. Incompressible medium flow path 9 (9a, 9b, 9c, 9d) communicating, the first medium storage tank 8, the second medium storage tank 10, 11, 12, and the incompressible medium flow path 9 are incompressible. Filled with medium 17. The openings of the second medium storage tanks 10, 11, 12 are covered and closed by elastically deformable diaphragm members 13, 14, 15. One end of the tube 4 is connected to the first medium storage tank 8 by a tube connector 16, and the other end of the tube 4 is connected to a syringe pump (not shown).

Although the size of the incompressible medium container 1 is not particularly limited, the present invention exhibits an excellent effect particularly when handling a very small amount of liquid. The depth is in the range of 1 μm to several mm, and the volume of the first medium storage tank 8 and the second medium storage tanks 10, 11, and 12 is 0.1 μL to several mL.
The incompressible medium container 1 is preferably made of a material having a strength that does not deform other than the diaphragm members 13, 14, and 15 with respect to the pressure applied to the incompressible medium 17. For example, glass, metal, or hard polymer material is used. Examples of the polymer include polymethyl methacrylate (PMMA), polycarbonate, polystyrene, polysulfone, polyethersulfone, polyetherimide, polyarylate, polymethylpentene, and 1,3-cyclohexadiene polymer.

The incompressible medium 17 is preferably a liquid that is stable and inert for a long period of time, a gel, or a solid that is highly elastic and deformable. In addition, an elastic polymer such as silicone oil, polyethylene glycol, polymer gel, and polydimethylsiloxane (PDMS) can be used.
The diaphragm members 13, 14, and 15 are made of an elastic material, and a silicone rubber film or the like can be used. A material that has gas permeability and does not have permeability to the incompressible medium 17 is preferable because the operation of sealing the incompressible medium 17 becomes easy. For example, a polytetrafluoroethylene (PTFE) porous film or a PDMS film can be used.

When the incompressible medium 17 is not liquid but is a solid substance such as a high molecular weight polymer, the diaphragm members 13, 14, and 15 need not be used. Further, when the incompressible medium 17 is accommodated after the incompressible medium container 1 and the liquid container 2 described later are set, the diaphragm members 13, 14, and 15 are not necessarily used.
When the incompressible medium in the syringe is discharged by the syringe pump, the incompressible medium flows into the incompressible medium container 1 through the tube 4 and the pressure in the incompressible medium container 1 increases. Since the incompressible medium container 1 is made of a material having a strength that does not deform except for the diaphragm members 13, 14, 15 with respect to the applied pressure, the diaphragm members 13, 14, 15 bulges outward (deforms so as to protrude toward the outside of the second medium storage tank 10, 11, 12). FIG. 2 shows an enlarged view of the diaphragm member 14 in the cross-sectional view of FIG.

  Equal pressure is applied to the diaphragm members 13, 14, and 15 from the inside. If a diaphragm material having an appropriate elastic strength is selected, the internal pressure can be made sufficiently larger than the reaction force received from the outside of the diaphragm members 13, 14, 15, that is, from the diaphragm of the liquid container 2, and each diaphragm member 13, The difference in reaction force received by 14 and 15 can be ignored. In this case, the volume ratio of the amount swelled to the outside of each diaphragm member 13, 14, 15 does not depend on the flow path design of the liquid container 2 or the properties of the stored liquid, and the second medium storage tanks 10, 11, It can be controlled by a size of 12. Moreover, since the incompressible medium is used, the sum of the discharge amount of the syringe pump and the volume swelled outside the diaphragm members 13, 14, 15 is equal. That is, the flow rate flowing through the liquid container 2 can also be quantitatively controlled by the discharge amount of the syringe pump.

This feature is suitable for a POC analyzer that analyzes unknown measurement objects having different liquid properties such as viscosity and makes the liquid container 2 disposable for each measurement. Since the flow rate ratio does not depend on the flow path shape of the liquid container 2, the dimensional accuracy requirement at the time of manufacturing the liquid container 2 is eased.
In the above example, the syringe pump is used as the transformer means, but other means can also be used. For example, a pressing mechanism as shown in FIG. 3 can be used. That is, the elastic film 18 is installed on the first medium storage tank 8, the sealed container 20 is provided on the elastic film 18, and the inside of the sealed container 20 is filled with the incompressible medium 21. When the pin 19 is pushed down by the actuator, the elastic film 18 is pressed downward, and the incompressible medium 17 in the incompressible medium container 1 is transformed.

In FIG. 4, the top view of the liquid container 2 and BB 'sectional drawing of this top view are shown. The liquid container 2 has a plate-like structure, and liquid storage tanks 22, 23 and 24 are provided at positions corresponding to the second medium storage tanks 10, 11 and 12 of the incompressible medium storage body 1. Furthermore, it has the waste liquid storage tank 26 and the flow path 25 which connects the liquid storage tanks 22, 23, and 24 with the waste liquid storage tank 26.
Although the size of the liquid container 2 is not particularly limited, the present invention exhibits an excellent effect particularly when handling a small amount of liquid. For example, the flow path 25 has a width and depth in the range of 1 μm to several mm. The volume of the liquid storage tanks 22, 23, 24 ranges from 0.1 μL to several mL.

  The material of the liquid container 2 is preferably chemically inert to the liquid to be stored, and glass, metal, polymer, or the like can be used depending on the liquid. Examples of the polymer include rigid plastic such as polymethyl methacrylate (PMMA), polycarbonate, polystyrene, polysulfone, polyethersulfone, polyetherimide, polyarylate, polymethylpentene, and 1,3-cyclohexadiene polymer, Examples thereof include elastic polymers such as dimethylsiloxane (PDMS). When measuring the liquid in the liquid container 2 using an optical detection means such as a thermal lens, it is desirable to use a material that is transparent and does not absorb the measurement wavelength.

  The openings of the liquid storage tanks 22, 23, 24 are covered and sealed with the diaphragms 27, 28, 29, respectively. Depending on how the liquid storage tanks 22, 23, 24 are used, the diaphragms 27, 28, 29 are fixed to the liquid container 2 in advance, and when the liquid storage tanks 22, 23, 24 are put in use, they are pasted. May be fixed. The diaphragms 27, 28, and 29 may be in the form of a deformable sheet that has gas permeability and does not have permeability to the liquid stored in the liquid storage tanks 22, 23, and 24. PTFE porous membranes and porous membranes of various materials can be used. However, it is preferable to use a hydrophobic organic polymer or an inorganic material so that the liquid does not pass through the diaphragms 27, 28, and 29.

  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, and 1,3-cyclohexadiene polymer.

Cellulose acetate membranes may be used, but in the case of a reagent solution to which a surfactant is added, a highly hydrophobic membrane such as PTFE, silicone, polyethylene, etc. is more resistant to water permeation. Is preferable because it is large.
A membrane having a higher water pressure resistance can be fed at a higher pressure. However, the water pressure resistance of the membrane that can be used for the liquid container 2 in the present invention is 9 in the flow channel configuration as described in the section of the best mode for carrying out the invention. .8 kPa or more is preferable. The average pore diameter of the membrane is preferably from 0.1 μm to 5 μm. Considering that the smaller the pore diameter, the higher the water pressure resistance and the smaller the amount of permeated air, the most preferred is about 0.1 μm. The film thickness is preferably 25 to 300 μm.

  In addition, the liquid container 2 may be composed of one member or may be composed of a plurality of members. Examples of the latter include the following. The liquid container 2 is configured by bonding a pair of flat plate members, and at least one of the pair of flat plate members is provided with a groove on the plate surface, and the plate surface provided with the groove is attached inside. The flow path 25 is formed by combining them. The same applies to the incompressible medium container 1, in which a pair of flat plate members are bonded to each other, and the flow path 9 is formed by bonding the plate surfaces provided with the grooves on the inside. But you can.

  FIG. 5 shows the configuration of an analyzer equipped with the liquid delivery device of the present invention. The uncompressed raw medium container 1 shown in FIG. 1 is provided with three second medium storage tanks, and the diaphragm members of the respective second medium storage tanks are in contact with the three liquid storage tanks provided in the liquid container 2, respectively. Is arranged. A diaphragm is attached to the opening of each liquid storage tank so that each diaphragm member does not directly contact the liquid in the liquid storage tank. The liquid container 2 is placed on a stage 7 having a temperature control function. Depending on the type of chemical reaction (biochemical reaction), the temperature control function may not be provided.

  In order to measure a chemical reaction (biochemical reaction) in the flow path 25 of the liquid container 2, the laser emitting unit 5 is arranged at a position where the flow path 25 can be irradiated, and a signal from the irradiated flow path 25 is received. An optical detection unit 6 is provided at a position where it can be detected. In FIG. 5, the laser emission unit 5 and the optical detection unit 6 are arranged on the same axis facing each other across the flow path 25 as in the thermal lens signal measurement. There is no need. Further, the positional relationship between the laser emitting unit 5 and the optical detecting unit 6 may be reversed.

The analyzer provided with the liquid feeding device of the present invention is not limited to the one having the optical measuring means, and may be one using an electrical measuring means. For example, the chemical reaction (biochemical reaction) in the flow path 25 can be electrically measured by the liquid container 2 including the electrodes 35 and 36 in the flow path 25 as shown in FIG.
The analyzer includes an electric circuit for controlling and driving the function of the liquid feeding device. In addition, an actuator drive circuit and control circuit for driving the syringe pump 3 and controlling the flow rate, a heater drive circuit and temperature control circuit for controlling the stage temperature, a detection system control circuit for measuring the reaction, etc. An overall control CPU is provided for cooperative operation and control. As for the detection system, for example, in the case of thermal lens signal measurement, a light source drive circuit for laser irradiation, a light source modulation circuit for modulating laser intensity, a light detection circuit for detecting a signal from an optical sensor, and signal amplification A circuit, a signal processing circuit, and the like.
It is also possible to control each function of the analyzer from a personal computer or the like outside the analyzer and omit all or part of the control circuit in the analyzer. In this case, the analyzer is provided with a signal interface with a personal computer.

An operation procedure of analysis using this analyzer will be described with reference to plan views and cross-sectional views of (1) to (6) in FIG.
As shown in (1) of FIG. 6, first, the specimen is dropped into the liquid storage tank 23 of the liquid container 2, and the reagent solution for the specimen analysis reaction is dropped into the liquid storage tanks 22 and 24. In this example, an example of a two-reagent analysis reaction is shown, and different reagents enter the liquid storage tanks 22 and 24, respectively. The amount of dripping need not be exact. Alternatively, a dry reagent may be enclosed in advance in the liquid storage tanks 22, 23, and 24, and the lysis buffer may be dropped.

  Next, as shown in (2) of FIG. 6, the diaphragms 27, 28, and 29 are attached to the openings of the liquid storage tanks 22, 23, and 24, respectively, and sealed. Next, as shown in FIG. 6 (3), the microsyringe 33 is connected to the waste liquid tank 26 connected to the outlet of the channel 25, and air is injected into the channel 25. Then, the pressure in the flow path 25 of the liquid container 2 is increased, and the liquid stored in each liquid storage tank 22, 23, 24 is pushed toward the diaphragms 27, 28, 29. Since the diaphragms 27, 28, 29 are hydrophobic porous elastic films, only the gas existing between the diaphragms 27, 28, 29 and the liquid stored in the liquid storage tanks 22, 23, 24 is transmitted. Liquid does not leak out. Therefore, there is no gas between the diaphragms 27, 28, and 29 and the contained liquid.

Next, as shown in FIG. 6 (4), the incompressible medium container 1 is brought into contact with the liquid container 2 and fixed. At this time, the diaphragms 27, 28, 29 on the respective liquid storage tanks 22, 23, 24 of the liquid container 2 and the diaphragm members 13 provided in the respective second medium storage tanks 10, 11, 12 of the incompressible medium container 1. , 14 and 15 are arranged in contact with each other. Further, as shown in (5) of FIG. 6, when the non-compressible medium is discharged from the syringe by a syringe pump (not shown), the non-compressible medium flows into the non-compressible medium container 1 through the tube 4 and is not compressed. The pressure in the porous medium container 1 increases. Diaphragm members 13, 14, 15 provided in the respective second medium storage tanks 10, 11, 12 swell outward (deform so as to protrude toward the outside of the second medium storage tanks 10, 11, 12) and come into contact with each other. The diaphragms 27, 28, 29 on the liquid storage tanks 22, 23, 24 of the liquid container 2 are pressed downward. Therefore, the liquid in the liquid storage tanks 22, 23, 24 is pushed out into the flow path 25 and flows out into the flow path 25. This state is illustrated in an enlarged manner in (6) of FIG.
In FIG. 6 (5), the specimen and reagent that have flowed into the flow path 25 are mixed and reacted at a predetermined flow ratio determined by the design of the incompressible medium container 1. The total flow rate is also accurately controlled by the syringe pump. At a measurement point 32 located on the downstream side of the flow path 25, a laser is irradiated and quantitative measurement is performed by detecting a signal.

Next, the present invention will be described in more detail with reference to examples and reference examples.
[Example 1]
A PMMA microchip 1 shown in FIG. 7 was prepared as the incompressible medium container 1. The incompressible medium flow path 9 has a width of 200 μm and a depth of 100 μm. The incompressible medium flow path 9a has a length of 20 mm, and the incompressible medium flow paths 9e, 9f, and 9g all have a length of 25.7 mm. The diameters of the second medium storage tanks 10, 11, and 12 are 3 mm, 2.5 mm, and 5 mm, respectively. Silicone rubber membranes are bonded to the openings of the second medium storage tanks 10, 11 and 12, respectively. One end of a PEEK tube 4 is connected to the first medium storage tank 8 via a tube connector 16, and the other end of the tube 4 is connected to a syringe pump (not shown). The first medium storage tank 8, the second medium storage tanks 10, 11, 12 and the incompressible medium flow path 9 are filled with water and there are no residual bubbles.

  A PMMA microchip 2 shown in FIG. 8 was prepared as the liquid container 2. The channel 25 has a width of 200 μm and a depth of 100 μm. The length from the junction 30 to the junction 31 is 100 mm, and the length from the junction 31 to the measurement point 32 is 130 mm. The diameters of the liquid storage tanks 22, 23, and 24 are 3 mm, 2.5 mm, and 5 mm, respectively, and the depth is 2 mm.

  PTFE membranes 27, 28, and 29 were attached to the openings of the liquid storage tanks 22, 23, and 24, respectively, and a tube was connected to the waste liquid tank 26 at the outlet of the flow path 25 via a connector (none shown). . A syringe (not shown) was connected to the other end of the tube, and a suspension of polystyrene beads having a diameter of 1 μm was poured into the microchip 2 from this syringe. The suspension was injected until the bubbles in the liquid storage tanks 22, 23, and 24 were completely eliminated. Then, the syringe was removed from the tube, and a glass capillary with an inner diameter of 100 μm was connected instead.

  The microchip 1 in FIG. 7 and the microchip 2 in FIG. 8 were stacked and fixed on the stage as shown in FIG. The microchip 1 and the microchip 2 are the diaphragm members 13, 14, 15 on the second medium storage tanks 10, 11, 12 of the microchip 1, and the diaphragm 27 on the liquid storage tanks 22, 23, 24 of the microchip 2, 28 and 29 were placed in contact with each other.

When water is discharged from the syringe pump, the diaphragm members 13, 14, 15 on the microchip chip 1 are expanded outward, whereby the diaphragms 27, 28, 29 on the microchip 2 are pushed and the liquid storage tanks 22, 23, It deform | transforms so that it may protrude inside 24. For this reason, the bead suspension in the liquid storage tanks 22, 23, 24 flows into the flow path 25. The total amount of liquid fed from the liquid storage tanks 22, 23, 24 was calculated from the amount of movement of the meniscus of the glass capillary. FIG. 9 shows the results of measurement of the relationship between the discharge amount of the syringe pump and the liquid feed amount in the microchip 2. The discharge amount of the syringe pump and the liquid feed amount in the microchip 2 were proportional and matched with an accuracy of 5% or less.
Ten microchips 2 were prepared and the same experiment was performed. When the same liquid amount was discharged with a syringe pump, the variation in the liquid feeding amount in the microchip 2 was within 1.4%. When the flow of the suspension was observed with an optical microscope, there was no pulsation even when the discharge speed of the syringe pump was 0.1 μL / min, and the liquid flow was smooth.

[Example 2]
An incompressible medium container 1 and a liquid container 2 having the same shape as those used in Example 1 were prepared, and a fluorescent dye solution was sealed in the liquid storage tank 22 or the liquid storage tank 23 of the microchip 2. Similarly to Example 1, the microchip 1 and the microchip 2 were stacked, fixed on the stage, and water was discharged from the syringe pump connected to the microchip 1. The liquid flow was observed with an optical microscope at the confluence 30 or the confluence 31 of the flow path 25 of the microchip 2. The flow ratio from each liquid storage tank 22, 23, 24 can be measured because the liquid is visualized by the fluorescent dye. As a result, the flow rate ratio was always kept constant between the discharge amount of 0 μL and 25 μL of the syringe pump, which was the experimental range.
In addition, although dextran was further added to the liquid storage tank 23 and the viscosity of the liquid was changed, no change was observed in the flow rate ratio in the viscosity range of 0.9 mPa · s to 10 mPa · s.

Example 3
Next, an example applied to actual blood analysis will be described. A multi-lipid calibrator (manufactured by Wako Pure Chemical Industries, Ltd.) was used as a specimen. The dilution ratio was adjusted with distilled water, and samples having 6 levels of concentration between 0 to 320 mg / dL of total cholesterol were prepared. L type Wako CHO · H (manufactured by Wako Pure Chemical Industries, Ltd.) was used as the reagent solution. The incompressible medium container and the liquid container had the same shape as that used in Example 1.

In the liquid storage tank 23 of the microchip 2, 10 μL of the specimen further diluted to 1/10 concentration, 40 μL of the enzyme coloring liquid A in the liquid storage tank 22 and 15 μL of the enzyme coloring liquid B in the liquid storage tank 24 were dropped. Then, PTFE membranes 27, 28, and 29 were attached to the openings of the liquid storage tanks 22, 23, and 24, respectively, and sealed.
A syringe was connected to the waste liquid tank 26, and air was sent into the flow path 25 of the microchip 2 to increase the internal pressure. Bubbles between the PTFE membranes of the liquid storage tanks 22, 23, and 24 and the specimen and reagent solution were pushed out of the membranes, and no bubbles were present.

The microchip 1 and the microchip 2 were set in the analyzer as shown in FIG. The temperature control stage was temperature-controlled so that the inside of the flow path 25 of the microchip 2 was 37 ° C. Then, water was discharged from the syringe pump at a speed of 2.4 μL / min. At this flow rate, it takes about 1 minute for the reaction solution of the specimen and the enzyme coloring solution A to pass between the junction 30 and the junction 31, and the reaction solution to which the enzyme solution B is further added is the junction 31. It took about 1 minute to pass between and the measurement point 32. The thermal lens signal was measured at the measurement point 32 by using a thermal lens signal measuring device (excitation light wavelength 630 nm, modulation frequency 1 kHz, probe light wavelength 780 nm) that was uniquely configured.
The analysis results are shown in FIG. As can be seen from the graph, there was a high correlation between the cholesterol concentration in the adjusted diluted specimen and the output of the thermal lens signal obtained by the thermal lens measurement. From this result, it was revealed that blood biochemical analysis can be performed with the microchip.

  The liquid feeding device of the present invention can precisely feed a small amount of liquid, and since the liquid does not come into contact with parts other than the liquid container, it is suitable for a microanalysis device that dislikes contamination between samples. is there. When the liquid delivery device of the present invention is applied to an analyzer, various analyzes including POC analysis can be performed easily and accurately.

It is the top view and sectional drawing which show an example of the incompressible medium container of this invention. It is the fragmentary sectional view which expanded and showed the 2nd medium storage tank peripheral part of the incompressible medium container of FIG. It is sectional drawing explaining the modification of a medium displacement mechanism. It is the top view and sectional drawing which show an example of the liquid container of this invention. It is a block diagram which shows an example of the analyzer which used the liquid feeding apparatus of this invention. It is a schematic diagram which shows the procedure of liquid feeding operation by the liquid feeding apparatus of this invention. It is a top view which shows the flow-path shape of the incompressible medium container used in the Example. It is a top view which shows the flow-path shape of the liquid container used in the Example. It is a graph which shows the relationship between the discharge amount of a syringe pump, and the liquid feeding amount of the liquid in a liquid container. 4 is a graph showing the relationship between the total cholesterol concentration in standard serum measured in Example 2 and the thermal lens signal value. It is the top view and sectional drawing which show an example of the liquid container provided with the electrode for electrically measuring the chemical reaction (biochemical reaction) in a flow path.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Incompressible medium container 2 Liquid container 3 Syringe pump 4 Tube 5 Laser emission part 6 Optical detection part 7 Temperature control stage 8 First medium storage tank 9, 9a, 9b, 9c, 9d Incompressible medium flow path 10, 11, 12 Second medium storage tank 13, 14, 15 Diaphragm member 16 Tube connector 17 Incompressible medium 18 Elastic film 19 Pin 20 Sealed container 21 Incompressible medium 22, 23, 24 Liquid storage tank 25 Channel 26 Waste liquid tank 27, 28, 29 Diaphragm 30, 31 Junction point 32 Measurement point 33 Micro syringe 35, 36 Electrode

Claims (4)

By changing the volume of the liquid storage tank surrounded by the wall, the liquid filled in the liquid storage tank is sent to the flow path connected to the liquid storage tank, or to the flow path or the flow path. A liquid feeding device for sending a liquid stored in another connected storage tank to the liquid storage tank,
A liquid container having the liquid storage tank, and an incompressible medium container for storing an incompressible medium that transmits a pressure for causing a volume change of the liquid storage tank,
The liquid storage tank is composed of a diaphragm having elasticity that is deformable so that at least a part of the wall body protrudes toward the inside or the outside of the liquid storage tank,
The incompressible medium container includes a first medium storage tank that includes a transformer for generating the pressure and is filled with the incompressible medium, and is communicated with the first medium storage tank by an incompressible medium flow path. A second medium storage tank,
The liquid storage tank and the second medium storage tank are disposed adjacent to each other with the diaphragm interposed therebetween, and the diaphragm deforms following the pressure change applied to the incompressible medium by the transformer. A liquid feeding device characterized by being configured as described above. By changing the volume of the liquid storage tank surrounded by the wall, the liquid filled in the liquid storage tank is sent to the flow path connected to the liquid storage tank, or to the flow path or the flow path. A liquid feeding device for sending a liquid stored in another connected storage tank to the liquid storage tank,
A liquid container having the liquid storage tank, and an incompressible medium container for storing an incompressible medium that transmits a pressure for causing a volume change of the liquid storage tank,
The liquid storage tank is composed of a diaphragm having elasticity that is deformable so that at least a part of the wall body protrudes toward the inside or the outside of the liquid storage tank,
The incompressible medium container includes a first medium storage tank that includes a transformer for generating the pressure and is filled with the incompressible medium, and is communicated with the first medium storage tank by an incompressible medium flow path. A second medium storage tank,
The liquid storage tank and the second medium storage tank are disposed adjacent to each other across the diaphragm and a diaphragm member provided in the second medium storage tank so as to be in contact with the diaphragm. Following the pressure change applied to the incompressible medium by a transforming means, the diaphragm can be deformed so as to protrude toward the inside or the outside of the liquid storage tank, and the diaphragm can be changed according to the deformation of the diaphragm member. A liquid feeding device characterized by being deformed.   The liquid container is configured by bonding a pair of flat plate members, and at least one of the pair of flat plate members is provided with a groove on the plate surface, and the plate surface including the groove is attached inside. The liquid feeding device according to claim 1, wherein the flow path is formed by combining them.   The incompressible medium container is configured by bonding a pair of flat plate members, and at least one of the pair of flat plate members includes a groove on the plate surface, and the plate surface including the groove on the inside. The liquid feeding device according to claim 1, wherein the incompressible medium flow path is formed by bonding together.

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