JP2009139120A - Microtest chip, liquid quantitation method of microtest chip and test device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a microtest chip having a high degree of freedom in disposing flow channels and wasting less specimens and reagents for accurately quantitating and dividing specimens and reagents individually for respective flow channels with a simple operation, and a liquid quantitation method of the microtest chip and a test device. <P>SOLUTION: The microtest chip having a high degree of freedom in disposing the flow channels and wasting less specimens and reagents for accurately quantitating and dividing specimens and reagents individually for respective flow channels with a simple operation by delivering a liquid stored in a liquid storing part to a plurality of liquid quantitation parts via a plurality of branched flow channels and delivering downstream the liquid filling a plurality of the liquid quantitation parts, the liquid quantitation method of the microtest chip and the test device is provided. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a micro test chip, a liquid quantification method and a test apparatus for a micro test chip, and more particularly, a test and analysis of biological materials by gene amplification reaction, antigen-antibody reaction, etc., test and analysis of other chemical substances, organic synthesis, etc. The present invention relates to a micro test chip used for chemical synthesis of a target compound by the method, a liquid quantification method of a micro test chip, and a test apparatus.

  In recent years, by making full use of micromachine technology and ultrafine processing technology, devices and means (for example, pumps, valves, flow paths, sensors, etc.) for performing conventional sample preparation, chemical analysis, chemical synthesis, etc. have been miniaturized. An analysis chip (hereinafter referred to as a micro inspection chip) integrated on a chip has been developed (see, for example, Patent Document 1).

  This is also called μ-TAS (Micro Total Analysis System), bioreactor, Lab-on-chip, biochip, and it is used in the medical examination / diagnosis field, environmental measurement field, and agricultural production field. Application is expected. In particular, when complicated processes such as those found in genetic testing, skilled techniques, and operation of equipment are required, micro test chips that excel in automation, speed-up, and simplification are cost-effective and require sample volume. Because it enables analysis not only for the required time but also for any time and place, the benefits are great.

  In the micro test chip as described above, it is important to accurately quantify the specimen and the reagent used for the test. If the sample or reagent cannot be accurately quantified, the reaction and the detection result will be greatly affected.

  Therefore, in Patent Document 2, the liquid introduced into the first flow path is drawn into the third flow path connecting the first flow path and the second flow path by capillarity, After removing the liquid in the flow path, the liquid in the third flow path is sent to the second flow path, thereby forming a liquid droplet having a volume corresponding to the volume of the third flow path. A method is disclosed.

Patent Document 3 discloses a method of quantifying the amount of liquid by the volume of a flow path by using a centrifugal force generated by rotating the chip to move the liquid in the chip.
JP 2004-28589 A JP 2002-357616 A JP 2000-514928 gazette

  However, in the method proposed in Patent Document 2, three flow paths are necessary only for quantifying the liquid, and the structure of the micro test chip becomes complicated. Further, when switching between the step of removing the liquid in the first flow path and the step of feeding the liquid in the third flow path to the second flow path, the force of the liquid feed pump is switched, It is necessary to devise measures such as sealing the end of the flow path in the middle of the liquid feeding process, which complicates the operation of the micro inspection chip. Furthermore, it is necessary to remove all the liquid remaining in the first flow path after the liquid is stored in the third flow path, and there is a lot of waste of specimens and reagents.

  In addition, the method proposed in Patent Document 3 has a problem in that it cannot be individually quantified for each flow path because liquid feeding by centrifugal force is simultaneously performed in all flow paths. Furthermore, since it is necessary to arrange the flow path in the direction from the center of rotation to the outside, there is a problem that the degree of freedom of arrangement of the flow path is small.

  The present invention has been made in view of the above circumstances, and it is possible to divide a specimen, a reagent, and the like accurately and individually for each flow path with a simple operation, and has a high degree of freedom in arranging the flow path. It is an object of the present invention to provide a micro test chip with little waste of reagents and the like, a liquid quantification method of the micro test chip, and a test apparatus.

  The object of the present invention can be achieved by the following constitution.

1. A liquid reservoir for storing liquid;
A plurality of liquid quantification units for quantifying the liquid stored in the liquid storage unit;
In a micro test chip comprising a plurality of downstream flow channels communicating with each of the plurality of liquid quantification units,
A plurality of branch passages communicated with the liquid storage section and each of the plurality of liquid determination sections;
An air flow path for feeding liquid that is communicated with the liquid reservoir and injects air into the liquid reservoir;
A micro-inspection chip comprising a plurality of air flow channels for quantification that communicate with each of the plurality of liquid quantification units and inject air into the plurality of liquid quantification units.

2. The plurality of liquid quantification units and the plurality of quantification air flow paths are communicated with each other via a water repellent valve.
2. The micro test chip according to 1, wherein the plurality of liquid quantification units and the plurality of downstream flow paths are communicated with each other via a water repellent valve.

  3. 3. The micro test chip according to 1 or 2, wherein the plurality of branch channels are provided with a high resistance channel having a high channel resistance value at least in a part thereof.

In the liquid test | inspection method of the micro test | inspection chip using the micro test | inspection chip of any one of 4.1 thru | or 3,
The liquid stored in the liquid storage part is supplied to the plurality of branch flow paths by the air injected from the liquid supply air flow path into the liquid storage part, and the plurality of liquid quantification parts are transferred to the liquid. A liquid feeding process that is fully filled with
A quantification step of sending the liquid fully filled in the plurality of liquid quantification units to the plurality of downstream channels by air injected from the plurality of quantification air channels into the plurality of liquid quantification units; A liquid quantification method for a micro inspection chip, comprising:

  5). 5. The injection of air from the liquid supply air flow path and the quantitative air flow path is performed by a driving liquid injected into the micro test chip from the outside of the micro test chip. Liquid quantification method for micro test chip

  6. An inspection apparatus for quantifying a liquid by using the liquid quantification method for a micro inspection chip according to 6.4 or 5.

  According to the present invention, the liquid stored in the liquid storage unit is sent to the plurality of liquid quantification units via the plurality of branch channels, and the liquid fully filled in the plurality of liquid quantification units is sent downstream. This makes it possible to divide specimens and reagents, etc., accurately and individually for each flow path with a simple operation, with a high degree of freedom in the arrangement of flow paths, and less waste of specimens and reagents, A liquid quantification method and an inspection apparatus for a micro inspection chip can be provided.

  Hereinafter, the present invention will be described based on the illustrated embodiment, but the present invention is not limited to the embodiment. In the drawings, the same or equivalent parts are denoted by the same reference numerals, and redundant description is omitted.

  First, an inspection apparatus according to the present invention will be described with reference to FIG. FIG. 1 is a schematic diagram showing an example of an inspection apparatus according to the present invention.

  In FIG. 1, the inspection apparatus 1 includes a micro inspection chip 100, a micro pump unit 210, a heating / cooling unit 230, a detection unit 250, a drive control unit 270, and the like.

  The micro inspection chip 100 is equivalent to what is generally called an analysis chip, a microreactor chip, and the like. For example, the micro inspection chip 100 is made of resin, glass, silicon, ceramics, and the like. Is formed with a fine flow path having a level of several μm to several hundred μm. The size and shape of the micro inspection chip 100 is usually a plate shape having a length and width of several tens of mm and a thickness of several mm.

  Here, the micro test chip 100 is formed of a water-repellent resin material such as polypropylene, for example, and has a channel substrate 101 on the surface of which a groove-shaped channel for flowing a liquid such as a reagent or a specimen is formed. Suppose that it is composed of a top plate 103 that is bonded to the surface of the flow path substrate 101 on which the flow path is formed and functions as a groove-shaped flow path cover of the flow path substrate 101. Further, the top plate 103 is provided with a connection port between the micro pump unit 210 and the micro test chip 100.

  The micro pump unit 210 is a pump unit for performing liquid feeding in the micro test chip 100, and includes a micro pump 211, a chip connection unit 213, a driving liquid tank 215, a driving liquid supply unit 217, and the like. The micropump unit 210 includes one or a plurality of micropumps 211. The micropump 211 feeds the liquid in the micro test chip 100 by injecting or sucking the driving liquid 216 into the micro test chip 100. The micro pump will be described in detail with reference to FIG. The chip connection unit 213 connects the micropump 211 and the micro inspection chip 100.

  The driving liquid supply unit 217 supplies the driving liquid 216 from the driving liquid tank 215 to the micropump 211. The driving liquid tank 215 can be removed and replaced from the driving liquid supply unit 217 to replenish the driving liquid 216. One or a plurality of pumps are formed on the micropump 211, and the plurality of pumps can be driven independently or in conjunction with each other.

  The micro test chip 100 and the micro pump 211 are connected and communicated with each other by a chip connection unit 213, and the micro pump 211 is driven to be injected or sucked from the micro pump 211 to the micro test chip 100 via the chip connection unit 213. With the liquid 216, various reagents and specimens stored in a plurality of storage units in the micro test chip 100 are sent in the micro test chip 100.

  The heating / cooling unit 230 includes a cooling unit 231, a heating unit 233, and the like, and heats and cools a specimen, a reagent, a mixed solution thereof, and the like in order to promote and suppress a reaction in the micro test chip 100. The cooling unit 231 includes a Peltier element or the like. The heating unit 233 includes a heater or the like. Of course, the heating unit 233 may also be formed of a Peltier element.

  The detection unit 250 includes a light source 251 such as a light emitting diode (LED) or a laser, a light receiving element 253 such as a photodiode (PD), and the like, and a target substance contained in a generated liquid obtained by a reaction in the micro inspection chip 100. Is detected optically at the position of the detection region 255 on the micro inspection chip 100.

  The drive control unit 270 includes a microcomputer, a memory, and the like (not shown), and drives, controls, and detects each unit in the inspection apparatus 1.

  Next, an embodiment of a micro inspection chip according to the present invention will be described with reference to FIG. FIG. 2 is a schematic diagram showing the configuration of the embodiment of the micro inspection chip 100 according to the present invention.

  In FIG. 2, on the surface of the flow path substrate 101 of the micro test chip 100, there are a liquid supply air flow path 141, a liquid reservoir 113, a liquid inlet 113i, a first flow path 111a, and a second flow path 111b. Is formed. Here, an example is shown in which the liquid stored in the liquid storage unit 113 is divided into the first flow path 111a and the second flow path 111b and quantified. Splitting is also effective.

  The liquid supply air flow path 141 and the liquid storage portion 113 are communicated with each other via a connection flow path 145. As will be described later with reference to FIG. 3, the liquid 151 is injected into the liquid reservoir 113 from the liquid inlet 113 i. The liquid inlet 113i is sealed with a sealing member 113s such as an adhesive tape as necessary.

  The first channel 111a includes a first branch channel 119a, a first liquid metering unit 125a, a first downstream channel 131a, a first metering air channel 143a, and the like. The first branch channel 119a has a first high-resistance channel 117a in which, for example, the channel resistance is set high by narrowing the channel cross-sectional area. The first liquid quantitative unit 125a and the first quantitative air flow path 143a are communicated with each other by a water repellent valve 123a.

  Further, the first liquid quantification unit 125a and the first downstream flow path 131a are communicated with each other by a water repellent valve 127a. The first branch channel 119a and the first liquid metering unit 125a communicate with each other at the position of the water repellent valve 123a via the first narrow channel 121a. The water repellent valve will be described in detail with reference to FIG.

  The configuration of the second channel 111b is the same, and “first” in the configuration of the first channel 111a described above may be read as “second” and the suffix “a” may be read as “b”. The first branch flow path 119a and the second branch flow path 119b communicate with the liquid storage section 113 at the branch section 115.

  In the present embodiment, in order to divide and quantitate the liquid into different volumes, the first liquid quantification unit 125a and the second liquid quantification unit 125b have different volumes by changing the channel length. However, of course, the same volume may be used if it is determined to be the same volume. Moreover, the liquid storage part 113 needs to have a volume more than the sum of the volume of the 1st liquid fixed_quantity | quantitative_assay part 125a and the volume of the 2nd liquid fixed_quantity | quantitative_assay part 125b at least.

  At the upstream ends of the liquid feeding air flow channel 141, the first quantitative air flow channel 143a, and the second quantitative air flow channel 143b, communication ports P1 and P2 with the micropump 211 shown in FIG. And P3 are provided. When the micro test chip 100 is inserted into the test apparatus 1, the communication ports P 1, P 2 and P 3 are communicated with the micro pump 211, and the driving liquid 216 is injected from the micro pump 211. Each flow path is filled with air 153 before the liquid is injected into the liquid reservoir 113.

  Subsequently, the liquid determination method in the present embodiment will be described with reference to FIGS. FIG. 3 is a flowchart showing the liquid quantification method in the present embodiment, FIG. 4 shows the state of the flow path in the middle of the liquid feeding process described later, and FIG. 5 shows the flow at the end of the liquid feeding process. FIG. 6 is a schematic diagram showing the state of the flow path in the middle of the quantitative process described later.

  In FIG. 3, in step S101, a liquid 151 such as a reagent or a specimen is injected into the liquid reservoir 113i from the liquid injection port 113i using a pipette or the like (injection process). After the liquid injection is completed, the liquid injection port 113i is sealed with the sealing member 113s in step S111 (sealing process). The steps so far are steps performed by the micro inspection chip 100 alone.

  Next, in step S121, the micro test chip 100 is inserted into the test apparatus 1, and the communication ports P1, P2, and P3 are communicated with the micro pump 211. In step S131, the driving liquid 216 is injected from the micro pump 211 into the liquid supply air flow path 141 through the communication port P1. The liquid 151 stored in the liquid storage section 113 is pushed out to the branch section 115 side by the pressure P1A of the air 153 in the liquid supply air flow path 141 pressed by the driving liquid 216.

  The injection pressure P1L of the driving liquid 216 at this time is a pressure at which the liquid 151 cannot pass through the water repellent valves 123a, 123b, 127a, and 127b by the pressure P1A of the air 153 pushed by the driving liquid 216.

  The liquid 151 pushed out to the branching unit 115 side is divided into the first liquid quantifying unit 125a and the second liquid quantifying unit 125b via the first branch channel 119a and the second branch channel 119b. Liquid is fed (liquid feeding process).

  FIG. 4 shows a state during the liquid feeding process. The liquid 151 pushed by the air 153 is fed to the middle of the first liquid quantification unit 125a and the second liquid quantification unit 125b via the first branch channel 119a and the second branch channel 119b. Yes.

  FIG. 5 shows a state at the end of the liquid feeding process. The liquid 151 pushed by the air 153 is fully filled in the first liquid quantitative unit 125a and the second liquid quantitative unit 125b. Since the liquid 151 cannot pass through the water repellent valves 123a, 123b, 127a, and 127b, when the liquid 151 is fully filled in the first liquid metering unit 125a and the second liquid metering unit 125b, the liquid feed is in that state. Stopped. In a state where the liquid 151 is fully filled, the injection of the driving liquid 216 from the micropump 211 is stopped.

  Returning to FIG. 3, in step S141, the driving liquid 216 is injected from the micro pump 211 into the first quantitative air channel 143a and the second quantitative air channel 143b through the communication ports P2 and P3. The The injection pressures P2L and P3L of the driving liquid 216 at this time are set to pressures at which the liquid 151 can pass through the water repellent valves 127a and 127b by the pressures P2A and P3A of the air 153 pushed by the driving liquid 216.

  The first liquid quantification unit 125a and the second liquid quantification are determined by the pressures P2A and P3A of the air 153 in the first quantification air channel 143a and the second quantification air channel 143b pushed by the driving liquid 216. The liquid 151 fully filled in the portion 125b is sent to the first downstream channel 131a and the second downstream channel 131b via the water repellent valves 127a and 127b (quantitative process).

  At this time, the liquid 151 in the first branch flow path 119a and the second branch flow path 119b is stored by the air 153 in the first quantitative air flow path 143a and the second quantitative air flow path 143b. Since it is pushed back slightly to the part 113 side, it does not flow out into the first liquid quantifying part 125a and the second liquid quantifying part 125b.

  Therefore, the volume of the liquid 151 sent to the first downstream channel 131a and the second downstream channel 131b is equal to the volume of the first liquid metering unit 125a and the second liquid metering unit 125b, and is accurate. The liquid is quantified and sent.

  FIG. 6 shows a state in the middle of the quantitative process. The liquid 151 fully filled in the first liquid quantitative unit 125a and the second liquid quantitative unit 125b is pushed into the air 153 in the first quantitative air channel 143a and the second quantitative air channel 143b. Then, the liquid is fed to the first downstream channel 131a and the second downstream channel 131b via the water repellent valves 127a and 127b.

  As described above, in the present embodiment, air pressure is used for liquid feeding. As a means for giving air pressure, a pump that directly feeds air may be used, but here, a micropump 211 described in detail in FIG. 7 is used. The advantages will be described.

  In the present embodiment, the first liquid quantification unit 125a and the second liquid quantification unit 125b are in communication with each other via the first branch channel 119a and the second branch channel 119b. The liquid 151 is divided and sent. In such a case, due to variations in the damper component of the air 153 in the first quantitative air flow path 143a and in the second quantitative air flow path 143b, the first quantitative air flow path 143a An imbalance occurs in time series between the air pressure P2A and the air pressure P3A in the second quantitative air flow path 143b.

  As a result, the liquid 151 that should originally remain in the first branch flow path 119a and the second branch flow path 119b, together with the liquid 151 in the first liquid determination unit 125a and the second liquid determination unit 125b, A phenomenon that the liquid flows out to the first downstream flow path 131a and the second downstream flow path 131b occurs, which causes the quantitative property of the liquid 151 to deteriorate.

  Therefore, in this embodiment, the micropump 211 described in detail with reference to FIG. The liquid was fed by the pressure of the air 153 sandwiched between them.

  In this way, the amount of the air 153 to which the pressures P2A and P3A for feeding the liquid are applied is set to a constant fine amount of about the volume of the first quantitative air flow path 143a and the second quantitative air flow path 143b. Can be kept to a small amount. As a result, variation in the damper component of the air 153 in the first quantitative air flow path 143a and in the second quantitative air flow path 143b can be suppressed to be small, and accurate quantitative determination can be performed. .

  Furthermore, in the present embodiment, a first high resistance in which a part of the first branch channel 119a and the second branch channel 119b is set to have a high channel resistance by, for example, reducing the channel cross-sectional area. The channel 117a and the second high resistance channel 117b were used. As a result, even if some variation remains in the damper component of the air in the first quantitative air flow path 143a and the second quantitative air flow path 143b, the pressure between the air pressures P2A and P3A Time series imbalances can be mitigated from being transmitted via the first branch channel 119a and the second branch channel 119b.

  As a result, the liquid 151 that should remain in the first branch flow path 119a and the second branch flow path 119b, together with the liquid 151 in the first liquid quantification unit 125a and the second liquid quantification unit 125b, The phenomenon of flowing out into the downstream flow channel 131a and the second downstream flow channel 131b can be alleviated, and accurate quantification can be performed.

  Note that the channel resistance values of the first high-resistance channel 117a and the second high-resistance channel 117b are sufficiently higher than the channel resistance values of the first downstream channel 131a and the second downstream channel 131b. It is desirable to be high. For example, when it is necessary to make the variation of the liquid 151 within about 10%, the first high resistance channel 117a and the second high resistance channel 117b, the first downstream channel 131a, The ratio of the channel resistance value to the second downstream channel 131b is desirably 10: 1 or more.

  As described above, according to the present embodiment, the liquid 151 stored in the liquid storage unit 113 is changed into the first branch flow path 119a and the second flow path by the pressure of the air 153 in the liquid supply air flow path 141. The liquid is fed through the branch flow path 119b, and the first liquid quantification unit 125a and the second liquid quantification unit 125b are fully filled. Next, the liquid 151 fully filled in the first liquid quantitative unit 125a and the second liquid quantitative unit 125b is used as the air 153 in the first quantitative air channel 143a and the second quantitative air channel 143b. Liquid is fed to the first downstream channel 131a and the second downstream channel 131b.

  In this way, it is possible to divide specimens, reagents, etc., accurately and individually for each flow path with a simple operation, with a high degree of freedom in arranging the flow paths, and with little waste of specimens, reagents, etc. An inspection chip, a liquid determination method and an inspection apparatus for a micro inspection chip can be provided.

  Next, an example of the micropump 211 used in the above-described embodiment will be described with reference to FIG. As the micro pump 211, various types such as a check valve type pump provided with a check valve in an inflow / outflow hole of a valve chamber provided with an actuator can be used, but a piezoelectric pump using a piezoelectric element as a drive source is used. Is preferred. 7 is a schematic diagram showing an example of the configuration of the micropump 211. FIG. 7A is a cross-sectional view showing an example of a piezo pump, FIG. 7B is a top view thereof, and FIG. 7C. FIG. 6 is a cross-sectional view showing another example of a piezo pump.

  7A and 7B, the micropump 211 includes a substrate 402 on which a first liquid chamber 408, a first channel 406, a pressurizing chamber 405, a second channel 407, and a second liquid chamber 409 are formed. The upper substrate 401 stacked on the substrate 402, the vibration plate 403 stacked on the upper substrate 401, the piezoelectric element 404 stacked on the side of the vibration plate 403 facing the pressurizing chamber 405, and the driving of the piezoelectric element 404 A drive unit (not shown) is provided.

  The drive unit and the two electrodes on both surfaces of the piezoelectric element 404 are connected by wiring using a flexible cable or the like, and a drive voltage is applied to the piezoelectric element 404 by the drive circuit of the drive unit through the wiring. . The first liquid chamber 408, the first flow path 406, the pressurizing chamber 405, the second flow path 407, and the second liquid chamber 409 are filled with the driving liquid 216.

  As an example, a photosensitive glass substrate having a thickness of 500 μm is used as the substrate 402, and etching is performed until the depth reaches 100 μm, whereby the first liquid chamber 408, the first flow path 406, the pressurizing chamber 405, the second A flow path 407 and a second liquid chamber 409 are formed. The first channel 406 has a width of 25 μm and a length of 20 μm. The second channel 407 has a width of 25 μm and a length of 150 μm.

  By stacking the upper substrate 401, which is a glass substrate, on the substrate 402, the upper surfaces of the first liquid chamber 408, the first flow path 406, the second liquid chamber 409, and the second flow path 407 are formed. A portion of the upper substrate 401 that corresponds to the upper surface of the pressurizing chamber 405 is processed by etching or the like to penetrate therethrough.

  A vibration plate 403 made of thin glass having a thickness of 50 μm is laminated on the upper surface of the upper substrate 401, and a piezoelectric element 404 made of, for example, lead zirconate titanate (PZT) ceramic having a thickness of 50 μm is laminated thereon. It is affixed. Due to the drive voltage from the drive unit, the piezoelectric element 404 and the vibration plate 403 attached thereto are vibrated, whereby the volume of the pressurizing chamber 405 is increased or decreased.

  The first flow path 406 and the second flow path 407 have the same width and depth, and the length of the second flow path 407 is longer than that of the first flow path 406. Then, when the differential pressure increases, turbulent flow is generated at and around the entrance / exit of the flow path, and the flow path resistance increases. On the other hand, since the length of the flow path in the second flow path 407 is long, it tends to become a laminar flow even if the differential pressure increases, and the rate of change in flow path resistance with respect to the change in differential pressure is smaller than that in the first flow path 406. Become. That is, the relationship of the ease of liquid flow between the first channel 406 and the second channel 407 changes depending on the magnitude of the differential pressure. Utilizing this, the drive voltage waveform for the piezoelectric element 404 is controlled to perform liquid feeding.

  For example, the vibration plate 403 is quickly displaced inward of the pressurizing chamber 405 by the driving voltage for the piezoelectric element 404 to reduce the volume of the pressurizing chamber 405 while applying a large differential pressure, and then is removed from the pressurizing chamber 405. If the volume of the pressurizing chamber 405 is increased while slowly displacing the vibration plate 403 in the direction and applying a small differential pressure, the fluid moves from the pressurizing chamber 405 to the second liquid chamber 409 (B in FIG. 7A). Direction).

  Conversely, the diaphragm 403 is quickly displaced outward from the pressurizing chamber 405 to increase the volume of the pressurizing chamber 405 while applying a large differential pressure, and then the diaphragm 403 is slowly moved inward from the pressurizing chamber 405. When the volume of the pressurizing chamber 405 is decreased while being displaced and a small differential pressure is applied, the fluid is fed from the pressurizing chamber 405 toward the first liquid chamber 408 (A direction in FIG. 7A). .

  Note that the difference in the flow rate resistance change ratio with respect to the change in differential pressure in the first flow path 406 and the second flow path 407 is not necessarily due to the difference in the length of the flow path, but is based on other geometric differences. It may be a thing.

  According to the micropump 211 configured as described above, the liquid feeding direction and the liquid feeding speed of a desired fluid can be controlled by changing the driving voltage and frequency of the pump. Although not shown in FIGS. 7A and 7B, the first liquid chamber 408 is provided with a port connected to the driving liquid tank 215 shown in FIG. It plays the role and receives the driving liquid 216 from the driving liquid tank 215 at the port. The second liquid chamber 409 forms a flow path of the micropump unit 210 shown in FIG. 1, and has a chip connection part 213 shown in FIG. 1 at the tip thereof, and is connected to the micro test chip 100 shown in FIG.

  In FIG. 7C, the micropump 211 includes a silicon substrate 471, a piezoelectric element 404, a substrate 474, and a flexible wiring (not shown). The silicon substrate 471 is obtained by processing a silicon wafer into a predetermined shape by a photolithography technique, and by etching, a pressurizing chamber 405, a diaphragm 403, a first channel 406, a first liquid chamber 408, a second channel 407, A second liquid chamber 409 is formed. The pressurizing chamber 405, the first channel 406, the first liquid chamber 408, the second channel 407, and the second liquid chamber 409 are filled with the driving liquid 216.

  The substrate 474 is provided with a port 472 above the first liquid chamber 408 and a port 473 above the second liquid chamber 409. For example, when the micropump 211 is separate from the micro test chip 100 Can communicate with the pump connection of the micro test chip 100 via the port 473. For example, the micro pump 211 can be connected to the micro test chip 100 by superimposing the substrate 474 in which the ports 472 and 473 are perforated and the vicinity of the pump connection part of the micro test chip 100 on each other.

  Further, as described above, since the micropump 211 is obtained by processing a silicon wafer into a predetermined shape by photolithography technology, a plurality of micropumps 211 can be formed on one silicon substrate. . In this case, it is desirable that the driving liquid tank 215 is connected to the port 472 opposite to the port 473 connected to the micro test chip 100. When there are a plurality of micropumps 211, their ports 472 may be connected to a common drive fluid tank 215.

  The above-described micropump 211 is small in size, has a small dead volume due to piping from the micropump 211 to the micro inspection chip 100, etc., has a small pressure fluctuation, and can instantaneously and accurately control discharge pressure. Accurate liquid feed control at 270 is possible.

  In the embodiment of the micro test chip 100 according to the present invention, water repellent valves 123a, 123b, 127a, and 127b are provided at both ends of the first liquid quantifying unit 125a and the second liquid quantifying unit 125b. Here, the general structure and operation of the water repellent valve will be described with reference to FIG. FIG. 8 is a schematic diagram for explaining a general structure and operation of a water repellent valve. FIG. 8A shows a state where liquid feeding is blocked by the water repellent valve. FIG. Indicates a state in which liquid is fed beyond the water repellent valve.

  In FIG. 8A, the water repellent valve 501 is configured by a small-diameter liquid feed control passage 511. The liquid feed control passage 511 has a cross sectional area S1 (a cross sectional area of a cross section perpendicular to the liquid feeding direction) smaller than a cross sectional area S2 of the upstream flow path 521 and a cross sectional area S3 of the downstream flow path 523. It is a narrow channel.

  When the flow path wall 531 is formed of a water-repellent material such as a plastic resin, the liquid 541 filled in the upstream flow path 521 is controlled by a weak liquid supply pressure P1 (for example, about 3 kPa). The flow into the channel 511 and the passage to the downstream channel 523 are restricted by the difference in surface tension between the channel wall 531 at the boundary between the liquid feeding control channel 511 and the downstream channel 523.

  In FIG. 8B, when the liquid 541 flows out to the downstream channel 523, a liquid feed pressure P2 (for example, about 10 kPa) higher than a predetermined pressure is applied by a micropump (not shown), and thereby the surface tension is increased. Against this, the liquid 541 is pushed out from the liquid feeding control passage 511 to the downstream flow passage 523. After the liquid 541 flows out to the downstream channel 523, the liquid 541 moves to the downstream channel 523 without maintaining the liquid feeding pressure P required to push the tip of the liquid 541 to the downstream channel 523. It will flow.

  That is, the passage of the liquid 541 from the liquid supply control passage 511 to the previous passage is blocked until the liquid supply pressure in the positive direction from the upstream flow path 521 to the downstream flow path 523 reaches the predetermined pressure P2. The liquid 541 passes through the liquid supply control passage 511 by applying a liquid supply pressure equal to or higher than P2.

  As described above, the sizes of the upstream flow path 521, the downstream flow path 523, and the liquid supply control passage 511 are not particularly limited as long as the passage of the liquid 541 to the upstream flow path 521 and the downstream flow path 523 can be regulated. However, as an example, the liquid supply control passage 511 is formed so that the vertical and horizontal dimensions are about 25 μm × 25 μm with respect to the upstream flow path 521 and the downstream flow path 523 whose vertical and horizontal dimensions are 150 μm × 300 μm.

  Further, in order to increase the liquid supply pressure difference (P2-P1) for restricting the liquid 541 from passing through the liquid supply control path 511, the flow in the portion of the downstream channel 523 in contact with the liquid supply control path 511 is increased. As shown in FIG. 8, the wall surface 531 a of the road wall 531 desirably rises at a right angle with respect to the liquid feeding control passage 511.

  As described above, according to the present invention, the liquid stored in the liquid storage unit is supplied to the plurality of liquid quantification units via the plurality of branch channels, and the plurality of liquid quantification units are fully filled. The sample and reagent can be divided and accurately quantified individually for each flow path with a simple operation, and the degree of freedom in the arrangement of the flow path is high. It is possible to provide a micro test chip with little waste, a liquid quantification method of a micro test chip, and a test apparatus.

  The detailed configuration and detailed operation of each component constituting the micro test chip, the liquid quantification method of the micro test chip, and the test apparatus according to the present invention can be changed as appropriate without departing from the spirit of the present invention. is there.

It is a schematic diagram which shows an example of the test | inspection apparatus in this invention. It is a schematic diagram which shows the structure of embodiment of the micro test | inspection chip in this invention. It is a flowchart which shows the liquid fixed_quantity | assay method in embodiment. It is a schematic diagram which shows the state in the middle of the liquid feeding process in embodiment. It is a schematic diagram which shows the state in the end of the liquid feeding process in embodiment. It is a schematic diagram which shows the state in the middle of the fixed_quantity | assay process in embodiment. It is a schematic diagram which shows one example of a structure of a micropump. It is a schematic diagram for demonstrating the general structure and operation | movement of a water repellent valve.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Inspection apparatus 100 Micro test | inspection chip 101 Flow path board | substrate 103 Top plate 111a 1st flow path 111b 2nd flow path 113 Liquid storage part 113i Liquid injection port 113s Sealing member 115 Branch part 117a 1st high resistance flow path 117b Second high resistance flow path 119a First branch flow path 119b Second branch flow path 121a First narrow flow path 121b Second narrow flow path 123a Water repellent valve 123b Water repellent valve 125a First liquid metering section 125b First 2 liquid quantification unit 127a water repellent valve 127b water repellent valve 131a first downstream flow path 131b second downstream flow path 141 liquid feeding air flow path 143a first quantitative air flow path 143b second quantitative air Flow path 145 Connection flow path 151 Liquid 153 Air 210 Micro pump unit 211 Micro pump 213 Chip Connection portion 215 driving liquid tank 216 driving liquid 217 driving liquid supply section 230 of the heating and cooling units 231 cool section 233 heating unit 250 detecting unit 251 light source 253 light receiving element 255 detect regions 270 drive controller

Claims (6)

A liquid reservoir for storing liquid;
A plurality of liquid quantification units for quantifying the liquid stored in the liquid storage unit;
In a micro test chip comprising a plurality of downstream flow channels communicating with each of the plurality of liquid quantification units,
A plurality of branch passages communicated with the liquid storage section and each of the plurality of liquid determination sections;
An air flow path for feeding liquid that is communicated with the liquid reservoir and injects air into the liquid reservoir;
A micro-inspection chip comprising a plurality of air flow channels for quantification that communicate with each of the plurality of liquid quantification units and inject air into the plurality of liquid quantification units. The plurality of liquid quantification units and the plurality of quantification air flow paths are communicated with each other via a water repellent valve.
The micro test chip according to claim 1, wherein the plurality of liquid quantification units and the plurality of downstream flow paths are communicated with each other via a water repellent valve. The micro test chip according to claim 1, wherein the plurality of branch channels include a high resistance channel having a high channel resistance value at least in a part thereof. In the liquid determination method of the micro test chip using the micro test chip according to any one of claims 1 to 3,
The liquid stored in the liquid storage part is supplied to the plurality of branch flow paths by the air injected from the liquid supply air flow path into the liquid storage part, and the plurality of liquid quantification parts are transferred to the liquid. A liquid feeding process that is fully filled with
A quantification step of sending the liquid fully filled in the plurality of liquid quantification units to the plurality of downstream channels by air injected from the plurality of quantification air channels into the plurality of liquid quantification units; A liquid quantification method for a micro inspection chip, comprising: The injection of air from the liquid feeding air channel and the quantitative air channel is performed by a driving liquid injected into the micro test chip from the outside of the micro test chip. The liquid quantification method of the micro test | inspection chip of description. An inspection apparatus for quantifying a liquid using the liquid quantification method for a micro inspection chip according to claim 4 or 5.

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