JPWO2009022496A1 - Micro inspection chip and inspection device - Google Patents

Abstract

A plurality of liquids are introduced from the mixed liquid flow path into the amplification unit, mixed, heated in the amplification unit to amplify the reaction to obtain a product solution, and the amplification unit detects the target substance in the product solution. The liquid can be efficiently mixed and reacted, and the target substance can be detected from the resulting product liquid. In addition, since the interface between the liquid mixture and air is heated after being sent to the liquid tip retention part with a small cross-sectional area, there is little evaporation and deterioration of the product liquid due to heating, and micro inspection suitable for downsizing and cost reduction Chips and inspection devices can be provided.

Description

  The present invention relates to a micro test chip and a test apparatus, and more particularly to biological substance inspection / analysis by gene amplification reaction, antigen-antibody reaction, etc., inspection / analysis of other chemical substances, chemical synthesis of target compounds by organic synthesis, etc. The present invention relates to a micro inspection chip and an inspection apparatus used.

  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 integrated on the 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, skilled techniques, and equipment operations are required as seen in genetic testing, an analytical chip with excellent automation, high speed, and simplification is cost-effective and requires a small amount of sample. Because it enables analysis not only for the required time but also for any time and place, the benefits are great.

  In various types of analysis and inspection, importance is attached to the quantitativeness of analysis, the accuracy of analysis, and the economy of these analysis chips. For that purpose, it is a problem to establish a highly reliable liquid feeding system with a simple configuration, and there is a demand for a microfluidic control element having high accuracy and excellent reliability. The inventors of the present invention have already proposed a micropump system suitable for this and a control method thereof (for example, see Patent Documents 2 to 4).

  In the analysis chip as described above (hereinafter referred to as a micro test chip), it is desirable that a mixed reagent obtained by mixing a plurality of reagents and a sample can be reacted. Therefore, in the micro test chip, various mixing operations such as mixing of reagents and mixing of a reagent and a sample are required in one chip. In view of this, a method has been proposed in which two types of liquids are mixed from two liquid reservoirs through a Y-shaped channel and mixed into a long mixing channel, and the target substance is detected in the detection unit (for example, , See Patent Document 5).

In addition, a mixture of a reagent and a sample is poured into the reaction chamber, the temperature is circulated in the reaction chamber, the reaction is performed, the solution is sent to the detection chamber, and the presence of the target substance is detected through the window provided in the detection chamber. A method of performing a test has been proposed (see, for example, Patent Document 6).
JP 2004-28589 A JP 2001-322099 A JP 2004-108285 A JP 2004-270537 A US Pat. No. 6,743,399 Japanese Patent No. 3558294

  However, in the method proposed in Patent Document 5, the two types of liquids that flow into the mixing channel via the Y-shaped channel do not mix easily because they flow in a laminar flow in the mixing channel. Therefore, it is not mixed unless the length of the mixing channel is very long. For this reason, the mixing channel occupies a large area on the micro test chip, which is an obstacle to miniaturization and cost reduction of the micro test chip. Moreover, since it is necessary to send the liquid over a long distance, the load on the liquid-feeding micropump is also large.

  Further, even in the method proposed in Patent Document 6, since the reaction chamber and the detection chamber are separately provided, a large space is required on the micro inspection chip, which becomes an obstacle to downsizing and cost reduction of the micro inspection chip. .

  The present invention has been made in view of the above circumstances, it is possible to efficiently mix and react a plurality of liquids, detect a target substance from the product liquid obtained by the reaction, without deterioration of the product liquid, An object is to provide a micro inspection chip and an inspection apparatus suitable for downsizing and cost reduction.

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

1. A liquid mixture flow path for sending a liquid mixture of a plurality of liquids downstream,
In a micro test chip including an amplification unit that communicates with the downstream of the mixed liquid channel and obtains a product liquid by amplifying the reaction of the mixed liquid by heating,
A liquid tip retention part in which the tip of the mixed liquid stays downstream of the amplification part;
The flow path cross-sectional area at the center of the amplifying unit is S2,
When the flow path cross-sectional area of the liquid tip retention portion is S3,
S3 <S2
A micro inspection chip characterized by

  2. 2. The micro test chip according to 1, wherein the amplification unit has a detection region for detecting a target substance contained in the product solution.

3.1 Micro inspection chip according to 1 or 2;
A micropump for feeding the liquid mixture;
A heating unit for heating the mixed liquid to obtain the product liquid;
An inspection apparatus comprising: a detection unit that detects a target substance contained in the product liquid.

  According to the present invention, a plurality of liquids are allowed to flow from the mixed solution flow path to the amplification unit, mixed, heated in the amplification unit to amplify the reaction to obtain a product solution, and the amplification unit targets the target substance in the product solution. Therefore, it is possible to efficiently mix and react a plurality of liquids, and to detect the target substance from the resulting product liquid. In addition, since the interface between the liquid mixture and air is heated after being sent to the liquid tip retention part with a small cross-sectional area, there is little evaporation and deterioration of the product liquid due to heating, and micro inspection suitable for downsizing and cost reduction Chips and inspection devices can be provided.

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 the principal part of the micro test | inspection chip in this invention. It is a schematic diagram which shows the structure of the main flow paths of embodiment of this invention centering on the amplification part. It is a schematic diagram which shows the state into which the liquid mixture flowed into the amplification part from the liquid mixture flow path. It is a schematic diagram which shows the state by which the liquid mixture was sent to the liquid front end retention part. It is a schematic diagram which shows one example of a structure of a micropump. It is a schematic diagram for demonstrating the structure and operation | movement of a water repellent valve.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Test | inspection apparatus 100 Micro test | inspection chip 111 Amplification part 111a Amplification part wall surface 115 Liquid front end retention part 121 Reagent storage part 123 Reagent 125a Water repellent valve 125b Water repellent valve 127 Reagent inlet 129 Sealing member 131 Specimen storage part 133 Sample 135a Water repellent Valve 135b Water repellent valve 137 Specimen inlet 139 Sealing member 141 Mixed liquid flow path 151 Generated liquid discharge flow path 153 Water repellent valve 161 Mixed liquid, generated liquid 161a (Mixed liquid, generated liquid and air) interface 191 Sealed Sheet 210 Micropump unit 211 Micropump 213 Chip connection part 215 Drive liquid tank 216 Drive liquid 217 Drive 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 270 Drive control Part 401 Upper substrate 402 Substrate 403 Vibration plate 404 Piezoelectric element 405 Pressurization chamber 406 First flow path 407 Second flow path 408 First liquid chamber 409 Second liquid chamber 471 Silicon substrate 472 Port 473 Port 474 Substrate 501 Water repellent valve 511 Liquid feed control passage 521 Upstream flow path 523 Downstream flow path 531 Flow path wall 541 Liquid

  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 pump unit 210 performs liquid feeding in the micro test chip 100. The heating / cooling unit 230 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 detection unit 250 detects a target substance contained in a product solution obtained by a reaction in the micro test chip 100. The drive control unit 270 performs driving, control, detection, and the like of each unit in the inspection apparatus 1.

  The micropump unit 210 includes a micropump 211, a chip connection unit 213, a driving liquid tank 215, a driving liquid supply unit 217, and the like. The micro pump 211 feeds the driving liquid 216. The chip connection unit 213 connects the micropump 211 and the micro inspection chip 100. The driving liquid tank 215 supplies a driving liquid 216 for liquid feeding. 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 heating / cooling unit 230 includes a cooling unit 231 and a heating unit 233. 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 may also be composed of Peltier elements. 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 detects a target substance contained in a generated liquid obtained by a reaction in the micro inspection chip. Then, optical detection is performed at the position of the detection region 255 on the micro inspection chip 100.

  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. A fine channel having a thickness of several μm to several hundred μm is formed. 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.

  The micro test chip 100 and the micro pump 211 are connected to and communicated with each other through a chip connection unit 213, and the micro pump 211 is driven to drive various reagents and samples stored in a plurality of storage units in the micro test chip 100. However, the liquid is fed by the driving liquid 216 flowing into the micro test chip 100 from the micro pump 211 via the chip connection part 213.

  Next, the configuration of the main part of the micro test chip 100 according to the present invention will be described with reference to FIG. FIG. 2 is a schematic diagram showing a configuration of a main part of the micro inspection chip 100 according to the present invention. Here, the case where the plurality of liquids in the present invention are one reagent and one specimen is illustrated, but the present invention is not limited to this, and may be a reagent and a reagent or a plurality of reagents and a specimen.

  In FIG. 2, the micro test chip 100 includes a reagent storage unit 121, a sample storage unit 131, a mixed solution channel 141, an amplification unit 111, a liquid tip retention unit 115, a product solution discharge channel 151, a water repellent valve 153, and the like. Consists of roads. Each flow path is preferably made of a hydrophobic material such as plastic, or made hydrophobic by applying a hydrophobic coating to the inner surface of the flow path.

  The reagent storage unit 121 is provided with water repellent valves 125a and 125b on the upstream side and the downstream side, respectively. In addition, a reagent inlet 127 is provided in the reagent reservoir 121. By injecting an appropriate amount of the reagent 123 from the reagent injection port 127, the reagent 123 is quantified from the water repellent valves 125a to 125b and fully filled. After completion of filling of the reagent 123, the reagent inlet 127 is sealed with a sealing member 129 made of an adhesive sheet or the like.

  Similarly, in the sample reservoir 131, the sample 133 is quantified from the water-repellent valves 135a to 135b and is fully filled by injecting an appropriate amount of the sample 133 from the sample inlet 137. After completion of the filling of the specimen 133, the specimen injection port 137 is sealed with a sealing member 139 made of an adhesive sheet or the like.

  The reagent 123 and the sample 133 stored in the reagent storage unit 121 and the sample storage unit 131 are moved downstream by the driving liquid 216 sent from the micropump 211 located upstream of the reagent storage unit 121 and the sample storage unit 131. The liquid is sent. The reagent reservoir 121 and the specimen reservoir 131 function as a plurality of liquid reservoirs in the present invention.

  FIG. 2 shows a state in which the reagent 123 and the specimen 133 are fed halfway through the mixed liquid channel 141. Since the reagent 123 and the sample 133 are sent as a laminar flow, they are not mixed so much in the mixed liquid channel 141, but the reagent 123 and the sample 133 merged in the mixed liquid channel 141 are referred to as a mixed solution 161. Thereafter, the mixed solution 161 is sent from the mixed solution channel 141 to the amplification unit 111, mixed and heated in the amplification unit 111, and the reaction is promoted to become a product solution 161.

  Next, an embodiment of the present invention will be described with reference to FIGS. FIG. 3 is a schematic diagram showing the configuration of the main flow path of the embodiment of the present invention centering on the amplifying unit 111, and FIG. 3A shows the surface on which the flow path of the micro test chip 100 is formed ( FIG. 3B is a view showing the AA ′ cross section of FIG. 3A with the flow path face down. For convenience of description with reference to cross-sectional views, the view from the flow path surface side of FIG. 3 and subsequent figures shows that the portion downstream from the mixed liquid flow path 141 in FIG. It is shown with a degree of rotation.

  In FIG. 3A, the width of the mixed liquid channel 141 is W1. A wall surface 111 a of the amplifying unit 111 at a portion where the mixed liquid channel 141 and the amplifying unit 111 communicate with each other is a wall surface perpendicular to the mixed liquid channel 141. The amplification unit 111 has a substantially quadrangular shape with rounded corners, and the width near the center is W2. A taper is provided on the downstream side of the amplifying unit 111 so as to communicate with the liquid tip staying unit 115 having a narrow channel cross-sectional area. The width of the liquid tip retention portion 115 is W3. A product liquid discharge channel 151 having a water repellent valve 153 at the center is provided on the downstream side of the liquid tip retaining part 115.

  Near the center of the amplification unit 111, a detection region 255 for optically detecting a target substance contained in a product solution obtained by the reaction in the amplification unit 111 is provided. At least a portion of the detection region 255 of the amplification unit 111 and a sealing sheet 191 to be described later can transmit light.

  Furthermore, a heating unit 233 configured by a heater or the like illustrated by a broken line is provided on the channel surface side of the amplification unit 111 so as to avoid an optical path for detection by the detection unit 250. Although FIG. 1 shows an example in which heating by the heating unit 233 and optical detection by the detection unit 250 are performed at different positions on the micro inspection chip 100, in this embodiment, both heating and detection are amplified. This is performed by the unit 111.

  In FIG. 3B, the height of the mixed liquid channel 141 is H1. A wall surface 111 a of the amplifying unit 111 at a portion where the mixed liquid channel 141 and the amplifying unit 111 communicate with each other is a wall surface perpendicular to the mixed liquid channel 141. The amplification unit 111 has a substantially quadrangular shape with rounded corners, and the height near the center is H2. A taper is provided on the downstream side of the amplifying unit 111 so as to communicate with the liquid tip staying unit 115 having a narrow channel cross-sectional area. The height of the liquid tip retention part 115 is set to H3. A product liquid discharge channel 151 having a water repellent valve 153 at the center is provided on the downstream side of the liquid tip retaining part 115. Each flow path described above is sealed with a sealing sheet 191 from the side of the surface on which the flow paths on the micro test chip 100 are formed.

Here, if the cross-sectional area of the mixed liquid channel 141 is S1 (= W1 × H1) and the cross-sectional area near the center of the amplifier 111 is S2 (= W2 × H2),
S1 <S2
It is.

Further, when the cross-sectional area of the liquid tip staying portion 115 is S3 (= W3 × H3),
S3 <S2
It is.

As an example, when the width of the mixed liquid channel 141 is 0.2 mm and the height is 0.25 mm, the cross-sectional area S1 is 0.05 mm ² . Similarly, when the width of the amplification unit 111 is 2 mm and the height is 1 mm, the cross-sectional area S2 is 2 mm ² , and when the width of the liquid tip retention portion 115 is 0.7 mm and the height is 0.7 mm, the cross-sectional area is S3 = 0.49 mm ² , and the above-described relationship is established.

  Above and below the detection region 255 provided in the central portion of the amplification unit 111, a detection unit 250 for optically detecting a target substance contained in the product liquid obtained by the reaction in the amplification unit 111 is provided. . The detection unit 250 includes a light source 251 and a light receiving element 253 arranged above and below the micro inspection chip 100. This is a so-called transmission type configuration. Of course, a so-called reflective configuration in which the light source 251 and the light receiving element 253 are disposed on the same side of the micro inspection chip 100 may be used.

  Furthermore, a heating unit 233 configured by a heater or the like is provided on the flow path surface side of the amplification unit 111 of the micro inspection chip 100 so as to avoid the optical path for detection by the detection unit 250. Any transparent heater that does not affect the detection of the detection unit 250 may be disposed on the entire flow path surface side of the amplification unit 111.

  Subsequently, a state in which the mixed solution 161 of the reagent 123 and the specimen 133 flows into the amplification unit 111 from the mixed solution channel 141 will be described with reference to FIGS. 4 and 5. FIG. 4 is a schematic diagram showing a state in which the mixed solution 161 flows into the amplifying unit 111 from the mixed solution channel 141, and FIG. 4A is a view seen from the channel surface side of the micro test chip 100, and FIG. ) Is a view showing the AA ′ cross section of FIG.

  4 (a) and 4 (b), the mixed liquid channel 141 and the wall surface 111a of the amplifying unit 111 are vertically communicated, and the inner wall surface of the amplifying unit 111 is hydrophobic. The mixed liquid 161 that has flowed out spreads along the wall surface 111 a, is sent into the amplifying section 111 while discharging the air in the amplifying section 111 to the product liquid discharge flow path 151, and is mixed with the air at the tip of the mixed liquid 161. Convection occurs in the liquid mixture 161 due to the surface tension of the interface 161a, and mixing is promoted. FIGS. 4A and 4B schematically show a state in which the interface 161a at the tip of the mixed solution 161 advances along with the liquid feeding.

  FIG. 5 is a schematic diagram showing a state in which the interface 161a at the tip of the mixed solution 161 is fed to the liquid tip retention portion 115, and FIG. 5 (a) is a view as seen from the flow path surface side of the micro test chip 100. FIG. 5B is a view showing the AA ′ cross section of FIG.

  In FIG. 5A, the liquid mixture 161 is stopped in a state where the interface 161a with the air at the front end is sent to the liquid front end retention portion 115, and the liquid mixture 161 is heated by the heating unit 233 to amplify the reaction. As a result, the product liquid 161 is obtained. The target substance generated by the reaction contained in the product solution 161 is optically detected by the detection unit 250. Since the amplifying unit 111 is formed to have a sufficiently large width W2 and height H2, it is possible to obtain a sufficient amount of target substance for optical detection.

  In addition, since the interface 161a with the air at the tip of the product liquid 161 stops at the liquid tip retention part 115 having a small cross-sectional area S3 (= W3 × H3), the product liquid 161 evaporates from the interface 161a by heating. It is less likely to be oxidized and deteriorated or altered by air.

  In the present embodiment, the specimen and the reagent are fed by feeding the driving liquid by the micropump. However, it is not always necessary to use the liquid pump such as the micropump, and the air pump The sample or reagent may be directly fed using

  As described above, according to the present embodiment, the mixed solution 161 of the reagent 123 and the specimen 133 is caused to flow from the mixed solution channel 141 into the amplifying unit 111 having a wide cross-sectional area S2, and the heating unit 233 is used. By heating, the reagent 123 and the sample 133 can be efficiently mixed and reacted to obtain the product solution 161. Furthermore, since the amplification unit 111 detects the target substance in the product solution 161 by the detection unit 250, the target substance can be efficiently detected from the product solution 161 obtained by the reaction.

  Further, since the tip of the product liquid 161 is heated after being fed to the liquid tip retention portion 115 having a small cross-sectional area S3, the evaporation, deterioration, and alteration of the product liquid due to heating are small. Moreover, since all of the mixing, heating, reaction, and detection described above are performed by the amplifying unit 111, it is excellent in space saving and is suitable for downsizing and cost reduction of the micro inspection chip.

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

  6A and 6B, 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. .

  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.

  On the upper surface of the upper substrate 401, a vibration plate 403 made of thin glass having a thickness of 50 μm is laminated, 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. The piezoelectric element 404 and the vibration plate 403 attached thereto are vibrated by the driving voltage from the driving unit, 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 moves in the direction from the pressurizing chamber 405 to the second liquid chamber 409 (B in FIG. 6A). 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 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. 6A). .

  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. 6A and 6B, the first liquid chamber 408 is provided with a port connected to the driving liquid tank 215, and the first liquid chamber 408 plays the role of a “reservoir”. The driving liquid is supplied from the driving liquid tank 215 at the port. The second liquid chamber 409 forms a flow path of the micro pump unit 210, and there is a chip connection part 213 at the tip, which is connected to the micro test chip.

  In FIG. 6C, 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 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 micro test chip 100 according to the embodiment of the present invention, water repellent valves are provided upstream and downstream of the reagent reservoir 121 and the specimen reservoir 131 and at the center of the product liquid discharge channel 151. Here, the structure and operation of the water repellent valve will be described with reference to FIG. FIG. 7 is a schematic diagram for explaining the structure and operation of the water repellent valve. FIG. 7A shows a state in which liquid feeding is blocked by the water repellent valve, and FIG. Indicates that liquid is being pumped over the valve.

  In FIG. 7A, 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 hydrophobic material such as a plastic resin, the liquid 541 filled in the upstream flow path 521 flows into the liquid supply control path 511 and the liquid supply control path 511. And passage to the downstream flow path 523 are restricted by the difference in surface tension between the flow path wall 531 at the boundary between the flow path 523 and the downstream flow path 523.

  In FIG. 7B, when the liquid 541 flows out to the downstream channel 523, a liquid feed pressure P equal to or higher than a predetermined pressure is applied by a micropump (not shown), thereby resisting the surface tension. 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 flows.

  That is, 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 P, the passage of the liquid 541 from the liquid supply control path 511 is blocked, and the predetermined pressure The liquid 541 passes through the liquid supply control passage 511 when a liquid supply pressure equal to or higher than P is applied.

  When the flow path wall 531 is formed of a hydrophilic material such as glass, it is necessary to apply a water-repellent coating, for example, a fluorine-based coating, to at least the inner surface of the liquid feeding control passage 511.

  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.

  As described above, according to the present invention, a plurality of liquids are allowed to flow from the mixed solution flow path to the amplification unit, mixed, and heated in the amplification unit to amplify the reaction to obtain a product liquid, and the amplification unit Thus, the target substance in the product liquid is detected, so that a plurality of liquids can be efficiently mixed and reacted, and the target substance can be detected from the resulting product liquid. In addition, since the interface between the liquid mixture and air is heated after being sent to the liquid tip retention part with a small cross-sectional area, there is little evaporation and deterioration of the product liquid due to heating, and micro inspection suitable for downsizing and cost reduction Chips and inspection devices can be provided.

  The detailed configuration and detailed operation of each component constituting the micro inspection chip and the inspection apparatus according to the present invention can be changed as appropriate without departing from the spirit of the present invention.

Claims (3)

A liquid mixture flow path for sending a liquid mixture of a plurality of liquids downstream,
In a micro test chip including an amplification unit that communicates with the downstream of the mixed liquid channel and obtains a product liquid by amplifying the reaction of the mixed liquid by heating,
A liquid tip retention part in which the tip of the mixed liquid stays downstream of the amplification part;
The flow path cross-sectional area at the center of the amplifying unit is S2,
When the flow path cross-sectional area of the liquid tip retention portion is S3,
S3 <S2
A micro inspection chip characterized by The micro test chip according to claim 1, wherein the amplifying unit has a detection region for detecting a target substance contained in the product solution. The micro test chip according to claim 1 or 2,
A micropump for feeding the liquid mixture;
A heating unit for heating the mixed liquid to obtain the product liquid;
An inspection apparatus comprising: a detection unit that detects a target substance contained in the product liquid.