JP2009133719A - Microinspection chip, liquid quantifying method of microinspection chip and inspection device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a microinspection chip capable of dividing a specimen or reagent at every channel while individually accurately quantifying the same at every channel by simple operation, also enhanced in the degree of freedom of the arrangement of the channel and also reducing the waste of the specimen or reagent, a liquid quantifying method of the microinspection chip and an inspection device. <P>SOLUTION: The microinspection chip is constituted so as to be capable of dividing the specimen or reagent while individually accurately quantifying the same at every channel by simple operation by injecting a liquid from a liquid injection port and sucking the same from a downstream channel after the whole of a liquid quantifying part and the region up to the midway of a branch channel are filled with the liquid, also enhanced in the degree of freedom of the arrangement of the channel and also reduces the waste of the specimen or reagent. The liquid quantifying method of the microinspection chip and the inspection device are also disclosed. <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. First and second liquid quantification units for storing liquid;
A liquid inlet that communicates with the first liquid metering unit and the second liquid metering unit and injects the liquid into the first liquid metering unit and the second liquid metering unit;
A first downstream flow path that communicates downstream of the first liquid quantification unit and that feeds the liquid stored in the first liquid quantification unit downstream;
In a micro test chip comprising a second downstream channel that is communicated downstream of the second liquid quantification unit and sends the liquid stored in the second liquid quantification unit downstream.
A first branch channel having a first air opening hole at one end, and the other end communicating with a portion where the first liquid metering unit and the first downstream channel are communicated;
A second branch channel having a second air opening hole at one end and the other end communicating with a portion where the second liquid metering unit and the second downstream channel communicated with each other; A micro inspection chip characterized by that.

2. A first injection flow path for communicating the liquid injection port with the first liquid metering unit;
A second injection channel for communicating the liquid injection port with the second liquid metering unit;
A first air inflow path having a first air inflow hole at one end and the other end communicating with the first injection flow path;
2. The micro inspection chip according to 1, further comprising a second atmospheric inflow passage having a second atmospheric inflow hole at one end and the other end communicating with the second injection passage.

In the liquid quantification method of a micro test chip using the micro test chip described in 3.1,
An opening step of opening the first and second atmosphere opening holes;
An injection step of injecting the liquid from the liquid injection port to fully fill the first and second liquid quantification units and injecting the liquid halfway through the first and second branch flow paths;
A sealing step of sealing the first and second atmosphere opening holes;
And a suction step of sucking the liquid from the first and second downstream flow paths in a state where the liquid inlet is opened.

In the liquid quantification method of the micro test chip using the micro test chip described in 4.2,
A first opening step of opening the first and second atmosphere opening holes;
A first sealing step for sealing the first and second air inflow holes;
Injecting the liquid from the liquid injection port, the first and second injection flow paths and the first and second liquid quantification units are fully filled, and the first and second branch flow paths An injection step of injecting the liquid halfway through,
A second sealing step for sealing the first and second air opening holes;
A second opening step of opening the first and second air inlet holes;
And a suction step of sucking the liquid from the first and second downstream flow paths.

  5. An inspection apparatus for quantifying a liquid using the liquid quantification method for a micro inspection chip according to 5.3 or 4.

  According to the present invention, by injecting liquid from the liquid injection port, filling the liquid partway and the middle of the branch flow path, and then sucking from the downstream flow path, a sample, a reagent, and the like can be flowed with a simple operation. To provide a micro test chip, a micro test chip liquid quantification method, and a test apparatus that can be divided while accurately quantifying each path individually, have a high degree of freedom in arrangement of flow paths, and less waste of specimens and reagents. Can do.

  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 micropump unit 210 functions as liquid feed pumps SPa and SPb described later.

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

  In FIG. 2, a liquid inlet 113, a first channel 111a, and a second channel 111b are formed on the surface of the channel substrate 101 of the micro test chip 100. As shown in FIG. 2, the first flow path 111 a and the second flow path 111 b are formed symmetrically with respect to the liquid injection port 113.

  The first flow path 111a includes a first injection flow path 121a, a first liquid metering unit 123a, a first water repellent valve 125a, a first downstream flow path 127a, a first branch pipe 131a, a first It is composed of a branch flow path 133a and a first atmosphere opening hole 135a.

  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 atmosphere opening hole 135a and the second atmosphere opening hole 135b are sealed with a sealing member 141 such as an adhesive tape as necessary.

  A first detection area 261a is provided in the vicinity of the upstream end of the first downstream flow path 127a. When liquid is supplied, the presence or absence of liquid at this position is detected by a liquid detection unit (not shown) provided in the inspection apparatus 1. Is detected. Similarly, a second detection area 261b is provided for the second downstream flow path 127b, and the presence or absence of liquid at this position is detected. The liquid detection unit described above may be a general sensor such as a light transmission type or a light reflection type. In this case, the first detection area 261a and the second detection area 261b need to transmit light to be used.

  The first injection channel 121a and the second injection channel 121b are connected at one end under the liquid inlet 113, and divide the liquid injected from the liquid inlet 113 as will be described later. Liquid is sent downstream. The downstream end of the first injection channel 121a is communicated with the upstream end of the first liquid metering unit 123a, and the downstream end of the second injection channel 121b is communicated with the upstream end of the second liquid metering unit 123b. Yes.

  The first liquid quantification unit 123a and the second liquid quantification unit 123b are directly communicated under the liquid inlet 113 without using the first injection channel 121a and the second injection channel 121b. May be.

  The first liquid quantification unit 123a and the second liquid quantification unit 123b have the same predetermined volume, and contribute to the liquid quantification described later.

  The downstream end of the first liquid quantitative unit 123a is communicated with the upstream end of the first downstream flow path 127a via the first water repellent valve 125a. In addition, the downstream end of the first liquid metering unit 123a is also connected to the upstream end of the first branch flow path 133a via the first branch pipe 131a at the connection point with the first water repellent valve 125a. Yes. A first atmospheric opening hole 135a is provided at the downstream end of the first branch flow path 133a. It is assumed that each flow path is filled with air 153 before liquid injection.

  When the micro inspection chip 100 is inserted into the inspection apparatus 1, the first downstream flow path 127a is connected to the first liquid feeding pump SPa configured by the micro pump 211 and the like on the downstream side.

  The configuration of the second liquid quantification unit 123b is the same, and “first” in the configuration of the first flow path 111a described above may be read as “second”, and the suffix “a” may be read as “b”.

  Subsequently, the liquid determination method in the first embodiment will be described with reference to FIGS. 3 to 5. FIG. 3 is a flowchart showing the liquid quantification method in the first embodiment, FIG. 4 shows the state of the flow path in the injection step described later, and FIG. 5 shows the state of the flow path in the suction step described later. It is a schematic diagram.

  In FIG. 3, in step S101, the first atmosphere opening hole 135a and the second atmosphere opening hole 135b are opened (opening step). In step S111, a liquid 151 such as a reagent or a specimen is injected from the liquid inlet 113 using a pipette or the like. The injected liquid 151 is divided and injected into the first liquid quantification unit 123a and the second liquid quantification unit 123b through the first injection channel 121a and the second injection channel 121b.

  The liquid 151 is fully filled in the first injection channel 121a and the first liquid metering unit 123a, and the second injection channel 121b and the second liquid metering unit 123b, and the first branch channel 133a and the first liquid metering unit 123b are filled. It is injected until it flows into the second branch channel 133b (injection step). Whether or not the liquid 151 has flowed into the first branch flow path 133a and the second branch flow path 133b is confirmed by visual observation, for example.

  FIG. 4 shows a state where the injection process is completed. When the liquid 151 is fully filled in the first injection channel 121a and the first liquid metering portion 123a and reaches the connection point between the first water repellent valve 125a and the first branch pipe 131a, the first The liquid 151 cannot flow downstream beyond the first water repellent valve 125a because it is regulated by the water repellent valve 125a. The water repellent valve will be described in detail with reference to FIG.

  On the other hand, since the first branch flow path 133a side is opened to the outside by the first atmosphere opening hole 135a, the liquid 151 flows into the first branch flow path 133a via the first branch pipe 131a. To do. The same applies to the second flow path 111b side.

  The amount of the liquid 151 flowing into the first branch channel 133a and the amount of the liquid 151 flowing into the second branch channel 133b depend on the dimensional accuracy of each channel, the surface condition of the inner wall of the channel, etc. Since an error occurs, it is preferable that the volume of the first branch flow path 133a and the second branch flow path 133b has a margin for the error of the inflowing liquid 151.

  Returning to FIG. 3, in step S <b> 121, the first atmosphere opening hole 135 a and the second atmosphere opening hole 135 b are sealed with the sealing member 141 (sealing process). The steps so far are steps performed by the micro inspection chip 100 alone.

  Next, in step S131, the micro test chip 100 is inserted into the test apparatus 1, the first downstream flow path 127a is the first liquid feed pump SPa, and the second downstream flow path 127b is the second liquid feed pump. Connected to SPb. Further, the first detection area 261 a and the second detection area 261 b are set at detection positions of a liquid detection unit (not shown) provided in the inspection apparatus 1.

  In step S141, the first liquid feeding pump SPa connected to the first downstream flow path 127a is driven in a state where the liquid inlet 113 is opened to the outside, and the liquid in the first liquid metering unit 123a is driven. 151 is sucked and fed to the first downstream flow path 127a. In step S143, it is confirmed whether or not the liquid supply of the liquid 151 in the first liquid fixed amount unit 123a is completed. Confirmation of the completion of liquid feeding may be performed, for example, based on the presence or absence of the liquid 151 in the first detection area 261a.

  When the liquid feeding is not completed (step S143; No), steps S141 and S143 are repeated until the liquid feeding is completed. When the liquid feeding is completed (step S143; Yes), in step S145, the first liquid feeding pump SPa is stopped, the suction of the liquid 151 is stopped (suction process), and the series of operations is ended. The same applies to the second flow path 111b side.

  FIG. 5 shows a state where the suction process is completed. When the first liquid feed pump SPa is driven, the air 153 in the first water repellent valve 125a and the first downstream flow path 127a shown in FIG. 4 is sucked downstream. At this time, since the first atmosphere opening hole 135a is sealed in step S121, the first branch pipe 131a and the first branch are caused by the negative pressure caused by the air 153 in the first branch flow path 133a. The liquid 151 in the flow path 133a does not flow out to the first water repellent valve 125a side.

  On the other hand, the liquid 151 in the first liquid metering unit 123a flows through the first water repellent valve 125a because the air 153 flows in from the liquid inlet 113 because the liquid inlet 113 is opened to the outside. It flows out to the first downstream flow path 127a. The volume of the liquid 151 flowing out to the first downstream flow path 127a is equal to the sum of the volume of the first injection flow path 121a and the volume of the first liquid quantifying unit 123a, and the accurately quantified liquid 151 is sent. Can be liquid. The same applies to the second flow path 111b side.

  As described above, according to the first embodiment, the first branch pipe 131a and the first branch flow path 133a and the second branch pipe 131b and the second branch flow path 133b are within the volume range. Thus, even if the liquid 151 is roughly injected beyond the volume of the first liquid quantifying unit 123a and the second liquid quantifying unit 123b, the liquid 151 to be fed can be easily divided into two and the volume can be accurately determined. Therefore, the injection process of the liquid 151 can be simplified.

  Next, a second embodiment of the micro test chip according to the present invention will be described with reference to FIG. FIG. 6 is a schematic diagram showing the configuration of the second embodiment of the micro test chip 100 according to the present invention.

  In the first embodiment, one end of the first injection channel 121a and the second injection channel 121b is directly communicated under the liquid inlet 113, but in the second embodiment, The first injection channel 121a and the second injection channel 121b are communicated with each other via a liquid storage part 143 in which the liquid injected from the liquid injection port 113 is stored.

  Further, the first injection channel 121a communicates with a first atmospheric inlet channel 145a provided with a first atmospheric inlet hole 147a at one end, and the second injection channel 121b has a second channel at one end. The second air inflow passage 145b provided with the air inflow hole 147b is communicated.

  The first air inflow hole 147a and the second air inflow hole 147b are sealed with a sealing member 141 such as an adhesive tape as necessary. The liquid inlet 113 is also sealed with an inlet sealing member 149 such as an adhesive tape as necessary. It is assumed that each flow path is filled with air 153 before liquid injection. Other configurations of the second embodiment are the same as those of the first embodiment.

  Next, a liquid quantification method according to the second embodiment of the present invention will be described with reference to FIGS. FIG. 7 is a flowchart showing a liquid quantification method according to the second embodiment. FIG. 8 shows the state of the flow path in the injection step described later, and FIG. 9 shows the state of the flow path in the suction step described later. It is a schematic diagram.

  In FIG. 7, in step S101, the first atmosphere opening hole 135a and the second atmosphere opening hole 135b are opened (first opening step), and in step S103, the first atmosphere inflow hole 147a and the second atmosphere opening hole 135b are opened. The inflow hole 147b is sealed (first sealing step). In step S111, a liquid 151 such as a reagent or a specimen is injected from the liquid injection port 113 into the liquid storage unit 143 using a pipette or the like. The liquid 151 injected into the liquid storage unit 143 is injected into the first liquid quantification unit 123a and the second liquid quantification unit 123b via the first injection channel 121a and the second injection channel 121b.

  The liquid 151 is fully filled in the first injection channel 121a and the first liquid metering unit 123a, and the second injection channel 121b and the second liquid metering unit 123b, and the first branch channel 133a and the first liquid metering unit 123b are filled. It is injected until it flows into the second branch channel 133b (injection step). Whether or not the liquid 151 has flowed into the first branch flow path 133a and the second branch flow path 133b is confirmed by visual observation, for example.

  FIG. 8 shows a state where the injection process is completed. When the liquid 151 is fully filled in the first injection channel 121a and the first liquid metering portion 123a and reaches the connection point between the first water repellent valve 125a and the first branch pipe 131a, the first The liquid 151 cannot flow into the first water repellent valve 125a because it is regulated by the water repellent valve 125a. The water repellent valve will be described in detail with reference to FIG.

  In addition, since the first atmospheric inflow hole 147a is sealed in step S103, the liquid 151 is changed by the pressure of the air stored in the first atmospheric inflow hole 147a and the first atmospheric inflow passage 145a. 1 cannot flow into the air inlet passage 145a.

  On the other hand, since the first branch flow path 133a side is opened to the outside by the first atmosphere opening hole 135a, the liquid 151 flows into the first branch flow path 133a via the first branch pipe 131a. To do. The same applies to the second flow path 111b side.

  An error occurs between the amount of the liquid 151 flowing into the first branch flow path 133a and the amount of the liquid 151 flowing into the second branch flow path 133b due to the dimensional accuracy of each flow path. The volume of the branch flow path 133a and the second branch flow path 133b is preferably provided with a margin for the error of the liquid 151 that flows in.

  Returning to FIG. 7, in step S121, the first atmosphere opening hole 135a and the second atmosphere opening hole 135b are sealed with the sealing member 141, and in step S123, the liquid inlet 113 is the inlet sealing member. Sealed at 149. Steps S121 and S123 function as a second sealing step in the present invention. In step S125, the first atmospheric inflow hole 147a and the second atmospheric inflow hole 147b are opened (second opening process). The steps so far are steps performed by the micro inspection chip 100 alone.

  Next, in step S131, the micro test chip 100 is inserted into the test apparatus 1, the first downstream flow path 127a is the first liquid feed pump SPa, and the second downstream flow path 127b is the second liquid feed pump. Connected to SPb. Further, the first detection area 261 a and the second detection area 261 b are set at detection positions of a liquid detection unit (not shown) provided in the inspection apparatus 1.

  In step S141, the first liquid feed pump SPa connected to the first downstream flow path 127a is driven, and the liquid 151 in the first liquid quantitative unit 123a is sucked, and the first downstream flow path 127a. The liquid is sent to In step S143, it is confirmed whether or not the liquid supply of the liquid 151 in the first liquid fixed amount unit 123a is completed. Confirmation of the completion of liquid feeding may be performed, for example, based on the presence or absence of the liquid 151 in the first detection area 261a.

  When the liquid feeding is not completed (step S143; No), steps S141 and S143 are repeated until the liquid feeding is completed. When the liquid feeding is completed (step S143; Yes), in step S145, the first liquid feeding pump SPa is stopped, the suction of the liquid 151 is stopped (suction process), and the series of operations is finished. The same applies to the second flow path 111b side.

  FIG. 9 shows a state where the suction process is completed. When the first liquid feed pump SPa is driven, the air 153 in the first water repellent valve 125a and the first downstream flow path 127a shown in FIG. 8 is sucked downstream. At this time, since the first atmosphere opening hole 135a is sealed in step S121, the first branch pipe 131a and the first branch are caused by the negative pressure caused by the air 153 in the first branch flow path 133a. The liquid 151 in the flow path 133a does not flow out to the first water repellent valve 125a side.

  On the other hand, since the first air inflow hole 147a is opened to the outside in step S125, the liquid 151 in the first liquid quantifying unit 123a flows in the air 153 from the first air inflow hole 147a. It becomes easy to move and flows out to the first downstream flow path 127a through the first water repellent valve 125a. The volume of the liquid 151 flowing out to the first downstream flow path 127a is equal to the sum of the volume of the first injection flow path 121a and the volume of the first liquid quantifying unit 123a, and the accurately quantified liquid 151 is sent. Can be liquid.

  In addition, since the liquid inlet 113 is sealed in step S123, the liquid 151 stored in the liquid inlet 113 and the liquid reservoir 143 is not sent downstream. The same applies to the second flow path 111b side.

  The sealing of the liquid inlet 113 in step S123 described above is not essential. In this case, the liquid 151 stored in the liquid inlet 113 and the liquid storage unit 143 is divided and sent to the downstream together with the liquid 151 stored in the first liquid quantitative unit 123a and the second liquid quantitative unit 123b. Is done.

  As described above, according to the second embodiment, the first branch pipe 131a and the first branch flow path 133a and the second branch pipe 131b and the second branch flow path 133b are within the volume range. Thus, even if the liquid 151 is roughly injected beyond the volume of the first liquid quantifying unit 123a and the second liquid quantifying unit 123b, the liquid 151 to be fed can be easily divided into two and the volume can be accurately determined. Therefore, the injection process of the liquid 151 can be simplified.

  In the first and second embodiments described above, the first liquid quantitative unit 123a and the second liquid quantitative unit 123b have been described as having the same predetermined volume, but this is essential. Instead, the first liquid quantitative unit 123a and the second liquid quantitative unit 123b may have different predetermined volumes. In this case, the left-right symmetry with respect to the liquid inlet 113 of the 1st flow path 111a and the 2nd flow path 111b mentioned above is not essential. Further, the number of liquid quantification units is not necessarily two, and may be three or more.

  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. 10 is a schematic diagram showing an example of the configuration of the micropump 211. FIG. 10A is a cross-sectional view showing an example of a piezo pump, FIG. 10B is a top view thereof, and FIG. 10C. FIG. 6 is a cross-sectional view showing another example of a piezo pump.

  10A and 10B, 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. When 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 flows in the direction from the pressurizing chamber 405 to the second liquid chamber 409 (B in FIG. 10A). Direction).

  Conversely, the diaphragm 403 is quickly displaced outward from the pressurizing chamber 405 to increase the volume of the pressurizing chamber 405 while giving 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 to give a small differential pressure, the fluid is fed from the pressurizing chamber 405 toward the first liquid chamber 408 (A direction in FIG. 10A). .

  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. 10A and 10B, the first liquid chamber 408 is provided with a port connected to the driving liquid tank 215 shown in FIG. 1, and the first liquid chamber 408 is a “reservoir”. The drive fluid 216 is supplied from the drive fluid 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. 10C, 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.

  When the micropump 211 shown in FIG. 10 is used as the liquid feeding pump of the first and second embodiments, the driving liquid 216 inside the micropump 211 is not emptied by sucking air. At least the inside of the second liquid chamber 409 needs to be filled with an amount of driving liquid 216 corresponding to the air to be sucked.

In addition to the micropump 211 described above, a pump capable of directly sucking air may be used as the liquid feed pump of the first and second embodiments. For example,
1) The above-described micro pump 211 designed to drive air,
2) A pump that sends air while opening and closing the valve,
3) A pump that sends air by moving the syringe in the cylinder back and forth,
4) Rotary type pump,
Etc. are considered.

  In the first and second embodiments of the micro-inspection chip 100 according to the present invention, between the first liquid quantification unit 123a and the first downstream channel 127a, and between the second liquid quantification unit 123b and the second Water repellent valves 125a and 125b are provided between the downstream flow path 127b. Here, the general structure and operation of the water repellent valve will be described with reference to FIG. FIG. 11 is a schematic diagram for explaining a general structure and operation of a water repellent valve. FIG. 11A shows a state in which liquid feeding is blocked by the water repellent valve. Indicates a state in which liquid is fed beyond the water repellent valve.

  In FIG. 11A, 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.

  However, as in the present invention, when the upstream flow path 521 is connected to a flow path through which the liquid 541 flows more easily than the liquid supply control path 511, the liquid 541 does not flow into the liquid supply control path 511. Instead, it stops immediately before the liquid feed control passage 511 or in the middle of the upstream flow path 521.

  In FIG. 11B, when the liquid 541 flows out to the downstream channel 523, a liquid feeding pressure P2 (for example, about 10 kPa) higher than a predetermined pressure is applied by a micropump (not shown), thereby surface tension. 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. 11, 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 can be easily injected by injecting the liquid from the liquid inlet, filling the liquid partway and the middle of the branch channel, and then suctioning from the downstream channel. Specimens, reagents, etc. can be divided while accurately quantifying each sample and reagent by operation, and the flow rate of the flow path is highly flexible. Methods and inspection devices can be provided.

  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 1st Embodiment of the micro test | inspection chip in this invention. It is a flowchart which shows the liquid fixed_quantity | assay method in 1st Embodiment. It is a schematic diagram which shows the state of the flow path in the injection | pouring process in 1st Embodiment. It is a schematic diagram which shows the state of the flow path in the suction process in 1st Embodiment. It is a schematic diagram which shows the structure of 2nd Embodiment of the micro test | inspection chip in this invention. It is a flowchart which shows the liquid fixed_quantity | assay method in 2nd Embodiment. It is a schematic diagram which shows the state of the flow path in the injection | pouring process in 2nd Embodiment. It is a schematic diagram which shows the state of the flow path in the suction process in 2nd 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 105 Downstream flow path 111a 1st flow path 111b 2nd flow path 113 Liquid inlet 121a 1st injection flow path 121b 2nd injection flow path 123a 1st 1st liquid quantification part 123b 2nd liquid quantification part 125a 1st water repellent valve 125b 2nd water repellent valve 127a 1st downstream flow path 127b 2nd downstream flow path 131a 1st branch pipe 131b 2nd Branch pipe 133a First branch flow path 133b Second branch flow path 135a First atmosphere release hole 135b Second atmosphere release hole 141 Sealing member 143 Liquid storage section 145a First atmosphere inflow path 145b Second atmosphere Inflow passage 147a First air inflow hole 147b Second air inflow hole 149 Inlet sealing member 151 Liquid 153 Air 210 Micropump 211 Micro pump 213 Chip connection part 215 Driving liquid tank 216 Driving liquid 217 Driving liquid supply part 230 Heating / cooling unit 231 Cooling part 233 Heating part 250 Detection part 251 Light source 253 Light receiving element 255 Detection area 261a First detection area 261b First 2 detection area 270 Drive controller

Claims (5)

First and second liquid quantification units for storing liquid;
A liquid inlet that communicates with the first liquid metering unit and the second liquid metering unit and injects the liquid into the first liquid metering unit and the second liquid metering unit;
A first downstream flow path that communicates downstream of the first liquid quantification unit and that feeds the liquid stored in the first liquid quantification unit downstream;
In a micro test chip comprising a second downstream flow channel that communicates downstream of the second liquid quantification unit and sends the liquid stored in the second liquid quantification unit downstream.
A first branch channel having a first air opening hole at one end, and the other end communicating with a portion where the first liquid metering unit and the first downstream channel are communicated;
A second branch channel having a second air opening hole at one end and the other end communicating with a portion where the second liquid metering unit and the second downstream channel communicated with each other; A micro inspection chip characterized by that. A first injection flow path for communicating the liquid injection port with the first liquid metering unit;
A second injection channel for communicating the liquid injection port with the second liquid metering unit;
A first air inflow path having a first air inflow hole at one end and the other end communicating with the first injection flow path;
2. The micro test chip according to claim 1, further comprising: a second air inflow path having a second air inflow hole at one end and the other end communicating with the second injection flow path. . In the liquid quantification method of the micro test chip using the micro test chip according to claim 1,
An opening step of opening the first and second atmosphere opening holes;
An injection step of injecting the liquid from the liquid injection port to fully fill the first and second liquid quantification units and injecting the liquid halfway through the first and second branch flow paths;
A sealing step of sealing the first and second atmosphere opening holes;
And a suction step of sucking the liquid from the first and second downstream flow paths in a state where the liquid inlet is opened. In the liquid quantification method of the micro test | inspection chip using the micro test | inspection chip of Claim 2,
A first opening step of opening the first and second atmosphere opening holes;
A first sealing step for sealing the first and second air inflow holes;
Injecting the liquid from the liquid injection port, the first and second injection flow paths and the first and second liquid quantification units are fully filled, and the first and second branch flow paths An injection step of injecting the liquid halfway through,
A second sealing step for sealing the first and second air opening holes;
A second opening step of opening the first and second air inlet holes;
And a suction step for sucking the liquid from the first and second downstream flow paths. An inspection apparatus characterized in that liquid is quantified using the liquid quantification method of a micro inspection chip according to claim 3 or 4.

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