CN106834115B - Microdroplet digital PCR chip module - Google Patents
Microdroplet digital PCR chip module Download PDFInfo
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- CN106834115B CN106834115B CN201710106548.8A CN201710106548A CN106834115B CN 106834115 B CN106834115 B CN 106834115B CN 201710106548 A CN201710106548 A CN 201710106548A CN 106834115 B CN106834115 B CN 106834115B
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- sample
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- 238000007847 digital PCR Methods 0.000 title claims abstract description 19
- 239000007788 liquid Substances 0.000 claims abstract description 114
- 239000000839 emulsion Substances 0.000 claims abstract description 52
- 239000000758 substrate Substances 0.000 claims abstract description 44
- 239000012530 fluid Substances 0.000 claims abstract description 5
- 238000007789 sealing Methods 0.000 claims abstract description 3
- 238000003860 storage Methods 0.000 claims description 41
- 238000002347 injection Methods 0.000 claims description 15
- 239000007924 injection Substances 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 3
- 238000005070 sampling Methods 0.000 claims 3
- 238000000465 moulding Methods 0.000 abstract description 2
- 238000001746 injection moulding Methods 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 4
- 150000007523 nucleic acids Chemical class 0.000 description 3
- 102000039446 nucleic acids Human genes 0.000 description 3
- 108020004707 nucleic acids Proteins 0.000 description 3
- 238000012408 PCR amplification Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000002411 adverse Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229920001577 copolymer Polymers 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000009434 installation Methods 0.000 description 1
- 229940127554 medical product Drugs 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000010008 shearing Methods 0.000 description 1
Classifications
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0689—Sealing
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/02—Identification, exchange or storage of information
- B01L2300/025—Displaying results or values with integrated means
- B01L2300/027—Digital display, e.g. LCD, LED
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/06—Auxiliary integrated devices, integrated components
- B01L2300/0609—Holders integrated in container to position an object
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/12—Specific details about materials
- B01L2300/123—Flexible; Elastomeric
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/08—Regulating or influencing the flow resistance
- B01L2400/084—Passive control of flow resistance
- B01L2400/086—Passive control of flow resistance using baffles or other fixed flow obstructions
Abstract
The invention discloses a microdroplet type digital PCR chip module, which comprises a chip, wherein the chip comprises a chip substrate, a chip bottom plate and a plurality of groups of emulsion microdroplet generating units arranged on the chip substrate; the clamping device comprises a bottom plate and a chuck, the chuck is of a rectangular frame body structure, four side edges of the chuck enclose a rectangular inner cavity for placing the chip, the chip is arranged in the rectangular inner cavity of the chuck, one end of the chip is propped against the sliding block, the other end of the chip is propped against the baffle, and the spring is in a compressed state; the emulsion droplet generation unit comprises 4 liquid reservoirs and a network through groove, wherein the liquid reservoirs are arranged on the top surface of the chip substrate, the network through groove comprises an oil phase droplet generation groove communicated with the oil phase liquid reservoirs and a sample droplet generation groove communicated with the sample liquid reservoirs, and the network through groove forms a sealing pipeline for fluid flow after the chip substrate is attached to the chip substrate. The module can collect a large amount of emulsion microdroplets and has the characteristics of convenient molding and better chip fastening.
Description
Technical Field
The invention relates to the field of biological detection and analysis equipment, in particular to a microdroplet digital PCR chip module.
Background
Digital droplet PCR (ddPCR) is a new PCR technology in recent years. The microdroplet digital PCR system performs microdroplet processing on the sample prior to conventional PCR amplification, i.e., the reaction system containing the nucleic acid molecules is divided into thousands of nanoupdated microdroplets, each of which contains either no nucleic acid target molecule to be detected or one to several nucleic acid target molecules to be detected. After PCR amplification, each droplet is detected one by one, the droplet with fluorescent signal is interpreted as 1, the droplet without fluorescent signal is interpreted as 0, and the initial copy number or concentration of the target molecule can be obtained according to the Poisson distribution principle and the number and proportion of positive droplets.
The core module of the microdroplet digital PCR device is a chip module that includes microscale channels that control the flow, heat transfer, and mass transfer of tiny fluids (typically picoliters to nanoliters) to create tiny droplets.
At present, a droplet-type digital PCR instrument of the company Limited for life medical products of the United states of America is more widely used, and the specific structure of the droplet-type digital PCR instrument is as shown in Chinese patent invention (grant date CN 103429331B grant date 2016.09.28), the invention discloses a system for forming an emulsion that may include an instrument and a microfluidic chip received by the instrument. The instrument may apply pressure to the desired emulsion phase held by the chip to drive the formation and collection of the emulsion in the chip. The chip comprises a plurality of groups of emulsion generating components, wherein one group comprises an oil phase liquid storage tank, a sample liquid storage tank and an emulsion collecting tank, the liquid in the oil phase liquid storage tank and the liquid in the sample liquid storage tank which is not mixed with the oil phase liquid storage tank are driven by using gas to apply pressure, and the liquid in the sample liquid storage tank which is not mixed with the gas are respectively connected into the emulsion collecting tank through and along the liquid drop generator. However, it is difficult to collect enough emulsion droplets in one emulsion collection tank, and the difficulty of injection molding of the chip structure of the three-row liquid storage tank is high, and the manufacturing cost is high.
Disclosure of Invention
In order to solve the above problems, an object of the present invention is to provide a microdroplet digital PCR chip module which can collect a large amount of emulsion microdroplets and has the characteristics of convenient molding and good chip fastening.
The micro-drop type digital PCR chip module comprises a chip, wherein the chip comprises a chip substrate, a chip bottom plate attached to the bottom surface of the chip substrate and a plurality of groups of emulsion micro-drop generating units arranged on the chip substrate; the chip clamping device comprises a bottom plate and a chuck;
the chuck is of a rectangular frame structure, and four side edges of the chuck enclose a rectangular inner cavity for placing a chip; two opposite long sides of the chuck are a first side and a second side respectively, two opposite wide sides of the chuck are a transverse plate and a baffle plate respectively, a chuck lug is arranged on the bottom surface of the transverse plate, one end of the baffle plate is hinged with the first side, and the other end of the baffle plate is clamped with the second side;
the chuck is inserted into the other side of the bottom plate groove through a chuck lug, and a spring is arranged between the slider and the chuck lug;
a chip is arranged in the rectangular inner cavity of the chuck, one end of the chip is propped against the sliding block, the other end of the chip is propped against the baffle, and the spring is in a compressed state;
the emulsion droplet generation unit comprises 4 liquid reservoirs arranged on the top surface of the chip substrate and network through grooves formed in the bottom surface of the chip substrate and communicated with the liquid reservoirs, the 4 liquid reservoirs are distributed on two sides of the chip substrate in two rows, and the 4 liquid reservoirs are an oil phase liquid reservoir, a sample liquid reservoir, a first collecting liquid reservoir and a second collecting liquid reservoir respectively; the network through tank comprises an oil phase droplet generation tank communicated with the oil phase liquid storage tank and a sample droplet generation tank communicated with the sample liquid storage tank, the oil phase droplet generation tank and the sample droplet generation tank are intersected in the emulsion generation tank, the outlet of the emulsion generation tank is divided into two paths, one path is communicated with the first collecting liquid storage tank, and the other path is communicated with the second collecting liquid storage tank; the network through groove forms a sealing pipeline for fluid flow after the chip substrate is attached to the chip bottom plate.
As a preferable scheme, the bottom of the oil phase liquid storage tank is provided with an oil phase sample injection micropore, the bottom of the sample liquid storage tank is provided with a sample injection micropore, the bottom of the first collecting liquid storage tank is provided with a first collecting micropore, and the bottom of the second collecting liquid storage tank is provided with a second collecting micropore; the oil phase microdroplet generating tank is communicated with the oil phase sample injection micropore through the oil phase liquid inlet tank, the sample microdroplet generating tank is communicated with the sample injection micropore through the sample liquid inlet tank, and the emulsion generating tank is respectively communicated with the first collecting micropore and the second collecting micropore through the emulsion buffer tank; the depth of each through groove in the network through groove is the same, the depth is 10-40 um, the width of the oil phase droplet generation groove is 0.2-0.3 times of the oil phase liquid inlet groove, the width of the sample droplet generation groove is 0.2-0.3 times of the sample liquid inlet groove, and the width of the emulsion generation groove is 0.2-0.3 times of the emulsion buffer groove.
As a preferable scheme, two oil phase droplet generation tanks are arranged, and the two oil phase droplet generation tanks are respectively communicated with the oil phase sample injection micropore through the oil phase liquid inlet tank. The structure of the two oil phase liquid flows formed by the two oil phase liquid inlet tanks for shearing and extruding the sample liquid flow to generate the micro-droplets is beneficial to the water-in-oil process, and emulsion micro-droplets are easier to obtain.
As a preferable scheme, the oil phase sample injection micropore and the sample injection micropore are respectively provided with a filter column group.
As a preferable scheme, 4 groups of emulsion droplet generation units are arranged on the chip substrate; the 4 groups of emulsion droplet generation units are sequentially and uniformly distributed along the length direction of the chip substrate.
As a preferable scheme, the bottom surface of the chip substrate is provided with a recess, and the top surface of the chip substrate is provided with a protrusion corresponding to the recess.
Preferably, the chip further comprises a cover plate arranged on the chip, and vent holes for injecting gas into the oil phase liquid storage tank and the sample liquid storage tank in the chip are arranged on the cover plate.
As a preferable scheme, the cover plate is provided with a positioning hole, the chuck is provided with a positioning column, and the positioning column penetrates into the positioning hole.
As a preferable scheme, the inner sides of the first side edge and the second side edge of the chuck are provided with mounting grooves matched with the chip, and the transverse plate of the chuck is provided with arc-shaped positioning grooves matched with the liquid storage tank.
Preferably, the chuck is fixed with the bottom plate through screws.
The invention has the advantages that:
1, because the microdroplet digital PCR device has higher requirement on compactness, the vibration and micro-shaking of the chip can generate adverse effects on the generation of micro liquid drops, and the clamping device mainly adopts a unilateral clamping mode, wherein the baffle, the sliding block and the spring can clamp the chip in the chuck.
2, two collecting liquid reservoirs are arranged in the emulsion droplet generation unit, and a large amount of emulsion can be collected by the first collecting liquid reservoir and the second collecting liquid reservoir compared with one collecting liquid reservoir.
3, the chip is mainly formed by injection molding, the manufacture of a die is key in the injection molding process, and for compact parts, the die core at the center of the die is required to have higher accuracy; in the prior art, the die core structure of the die for manufacturing the three rows of chips for distributing the liquid storage tanks is extremely complex, the processing difficulty is very high, and the manufacturing cost is very high; according to the invention, the three-row distributed liquid storage tank chips are transformed into the two-row distributed liquid storage tank chips, so that the processing difficulty of a die and a die core is reduced, and the injection molding of the cycloolefin copolymer (COC) chip is particularly facilitated.
Drawings
FIG. 1 is a schematic diagram of the structure of the present invention;
FIG. 2 is an exploded view of FIG. 1;
FIG. 3 is a schematic view of the chuck of FIG. 2;
FIG. 4 is an exploded view of the chip of FIG. 2;
FIG. 5 is a schematic diagram of the chip substrate of FIG. 4;
FIG. 6 is an enlarged view of section I of FIG. 5;
FIG. 7 is a front view of FIG. 5;
FIG. 8 is a cross-sectional view A-A of FIG. 7;
FIG. 9 is a cross-sectional view B-B of FIG. 7;
the reference numerals of the components in the drawings are as follows: chip 1 (including chip substrate 11, recess 11a, chip bottom plate 12, projection 12a, emulsion droplet generation unit 13, network through groove 130, oil phase liquid reservoir 131a, sample liquid reservoir 132a, first collection liquid reservoir 133a, second collection liquid reservoir 134a, oil phase liquid inlet tank 135, oil phase droplet generation tank 135a, sample liquid inlet tank 136, sample droplet generation tank 136a, emulsion buffer tank 137, emulsion generation tank 137a, filtration column group 138), clamping device 2 (including bottom plate 21, bottom plate groove 21.1, slider 21.2, chuck 22, rectangular inner cavity 22.1, first side 22.2, second side 22.3, cross plate 22.4, baffle 22.5, chuck bump 22.6, positioning column 22.7, mounting groove 22.8, arc positioning groove 22.9), spring 3,4 (including vent 41, positioning hole 42), and cover plate 5).
Detailed Description
For a better understanding of the present invention, the following detailed description of the invention will be given with reference to the accompanying drawings and specific examples.
Referring to fig. 1 and 2, a microdroplet digital PCR chip module includes a chip 1 and a clamping device 2 for the chip 1; as shown in fig. 4, the chip 1 comprises a chip substrate 11, a chip base plate 12 attached to the bottom surface of the chip substrate 11, and 4 groups of emulsion droplet generation units 13, wherein the 4 groups of emulsion droplet generation units 13 are arranged on the chip substrate 11 and are sequentially and uniformly distributed along the length direction of the chip substrate 11; as shown in fig. 1 and 2, the clamping device 2 includes a base plate 21 and a chuck 22, the bottom surface of the chuck 22 is attached to the base plate 21, and the chuck 22 is fixed to the base plate 21 by screws 5.
As shown in fig. 2, the chuck 22 has a rectangular frame structure, four side edges of the chuck 22 and a rectangular inner cavity 22.1 for placing the chip 1 are formed; two opposite long sides of the chuck 22 are a first side 22.2 and a second side 22.3 respectively, two opposite wide sides of the chuck 22 are a transverse plate 22.4 and a baffle plate 22.5 respectively, one end of the baffle plate 22.5 is hinged with the first side 22.2, and the other end of the baffle plate 22.5 is clamped with the second side 22.3; the inner sides of the first side 22.2 and the second side 22.3 of the chuck 22 are provided with mounting grooves 22.8 matched with the chip 1; the transverse plate 22.4 of the chuck 22 is provided with an arc-shaped positioning groove 22.9 matched with the liquid storage pool.
As shown in fig. 2, a bottom plate groove 21.1 is formed in the bottom plate 21, a sliding block 21.2 is arranged on one side in the bottom plate groove 21.1, as shown in fig. 3, a chuck lug 22.6 is arranged on the bottom surface of a transverse plate 22.4 of the chuck 22, the chuck lug 22.6 is inserted into the other side of the bottom plate groove 21.1, and a spring 3 is arranged between the sliding block 21.2 and the chuck lug 22.6;
a chip 1 is arranged in the rectangular inner cavity 22.1 of the chuck 22, one end of the chip 1 is propped against the sliding block 21.2, the other end of the chip 1 is propped against the baffle 22.5, and the spring 3 is in a compressed state; at this time, the chip 1 is clamped by the clamping device 2.
Referring to fig. 4 and 5, the emulsion droplet generating unit 13 includes 4 liquid reservoirs disposed on the top surface of the chip substrate 11 and a network through slot 130 formed on the bottom surface of the chip substrate 11 and communicating with the liquid reservoirs, as shown in fig. 7, the 4 liquid reservoirs are distributed on two sides of the chip substrate 11 in two rows, and the 4 liquid reservoirs are respectively an oil phase liquid reservoir 131, a sample liquid reservoir 132, a first collecting liquid reservoir 133 and a second collecting liquid reservoir 134; as shown in fig. 7 to 9, the bottom of the oil phase liquid storage pool 131 is provided with an oil phase sample injection micropore 131a, the bottom of the sample liquid storage pool 132 is provided with a sample injection micropore 132a, the bottom of the first collecting liquid storage pool 133 is provided with a first collecting micropore 133a, and the bottom of the second collecting liquid storage pool 134 is provided with a second collecting micropore 134a;
referring to fig. 5 to 6, the network through tank 130 includes two oil phase droplet generation tanks 135a and a sample droplet generation tank 136a, the two oil phase droplet generation tanks 135a are respectively communicated with the oil phase sample injection micro-holes 131a through the oil phase liquid inlet tanks 135, and the two oil phase liquid flows are sheared and extruded to form a sample liquid flow, so that emulsion droplets can be obtained more easily by the structure; the sample droplet generation well 136a communicates with the sample introduction microwell 132a through the sample inlet well 136; the oil phase droplet generation tank 135a and the sample droplet generation tank 136a are intersected in an emulsion generation tank 137a, the emulsion generation tank 137a is divided into two paths after passing through an emulsion buffer tank 137, one path is communicated with the first collecting liquid storage tank 133 through the emulsion buffer tank 137, and the other path is communicated with the second collecting liquid storage tank 134 through the emulsion buffer tank 137; the depth of each through groove in the network through groove 130 is the same and is 10um, the width of the oil phase droplet generation groove 135a is 0.2 times of the oil phase liquid inlet groove 135, the width of the sample droplet generation groove 136a is 0.2 times of the sample liquid inlet groove 136, and the width of the emulsion generation groove 137a is 0.2 times of the emulsion buffer groove 137. The oil phase sample introduction micropore 131a and the sample introduction micropore 132a are respectively provided with a filter column group 138. The network through groove 130 forms a sealed conduit for fluid flow after the chip substrate 11 is attached to the chip base plate 12.
As shown in fig. 4 and 5, the bottom surface of the chip substrate 11 is provided with a recess 11a, the top surface of the chip substrate 12 is provided with a protrusion 12a corresponding to the recess 11a, and the chip substrate 11 and the chip substrate 12 are bonded to each other by the recess 11a and the protrusion 12a so that the chip substrate 12 is tightly attached to the bottom surface of the chip substrate 11.
As shown in fig. 2, the microdroplet digital PCR chip module further includes a cover plate 4 disposed on the chip 1, and the cover plate 4 is provided with a vent 41 for injecting gas into the oil phase liquid reservoir 131 and the sample liquid reservoir 132 in the chip 1, where the gas injection is to apply air pressure into the oil phase liquid reservoir 131 and the sample liquid reservoir 132, so as to ensure that the oil phase in the oil phase liquid reservoir 131 and the sample in the sample liquid reservoir 132 circulate along the network pipeline. The cover plate 4 is further provided with a positioning hole 42, the chuck 22 is provided with a positioning column 22.7, the positioning column 22.7 penetrates into the positioning hole 42, and the positioning hole 42 and the positioning column 22.7 cooperate to enable the vent hole 41 on the cover plate 4 to correspond to the oil phase liquid storage pool 131 and the sample liquid storage pool 132 one by one, so that the situation that dislocation gas cannot be introduced is prevented.
The installation method of the embodiment comprises the following steps: as shown in fig. 2, the slider 21.2 is placed on one side of the bottom plate groove 21.1, the chuck 22 is placed on the bottom plate 21, the chuck lug 22.6 is inserted to the other side of the bottom plate groove 21.1, the spring 3 is arranged between the slider 21.2 and the chuck lug 22.6, the chuck 22 and the bottom plate 21 are fixed by the screw 5, the baffle 22.5 is lifted upwards, the chip 1 is inserted into the rectangular inner cavity 22.1 along the mounting groove 22.8 until one end of the chip 1 abuts against the slider 21.2, the spring 3 is forced to be in a compressed state, the baffle 22.5 is put down and clamped on the second side 22.3 of the chuck 22, and at the moment, the other end of the chip 1 abuts against the baffle 22.5, and the spring 3 is in a compressed state; at this time, the chip 1 is clamped by the clamping device 2. Then cover 4 is covered on chip 1 through the cooperation of locating hole 42 and reference column 22.7, and air vent 41 and oil phase liquid reservoir 131 and sample liquid reservoir 132 one by one on cover 4.
The microdroplet digital PCR chip module clamps the chip 1 through the clamping device 2 on one hand, on the other hand, 4 liquid reservoirs and the network through grooves 130 on the chip 1 form a group of emulsion microdroplet generating units 13, two liquid reservoirs for collecting emulsion are included in the 4 liquid reservoirs, so that a large amount of emulsion can be collected, the 4 liquid reservoirs are distributed on two sides of the chip substrate 11 in two rows, and the structure is convenient for injection molding of the template.
The above examples represent only 1 embodiment of the present invention, and the description thereof is more specific and detailed, but should not be construed as limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.
Claims (5)
1. The microdroplet type digital PCR chip module comprises a chip (1), wherein the chip (1) comprises a chip substrate (11), a chip bottom plate (12) attached to the bottom surface of the chip substrate (11) and a plurality of groups of emulsion microdroplet generation units (13) arranged on the chip substrate (11); the method is characterized in that: the chip clamping device comprises a clamping device (2) of the chip (1), wherein the clamping device (2) comprises a bottom plate (21) and a chuck (22);
the chuck (22) is of a rectangular frame structure, four side edges of the chuck (22) are surrounded, and a rectangular inner cavity (22.1) for placing the chip (1) is formed; two opposite long sides of the chuck (22) are a first side (22.2) and a second side (22.3) respectively, two opposite wide sides of the chuck (22) are a transverse plate (22.4) and a baffle plate (22.5) respectively, chuck convex blocks (22.6) are arranged on the bottom surface of the transverse plate (22.4), one end of the baffle plate (22.5) is hinged with the first side (22.2), and the other end of the baffle plate (22.5) is clamped with the second side (22.3);
a bottom plate groove (21.1) is formed in the bottom plate (21), a sliding block (21.2) is arranged on one side in the bottom plate groove (21.1), the chuck (22) is inserted into the other side of the bottom plate groove (21.1) through a chuck lug (22.6), and a spring (3) is arranged between the sliding block (21.2) and the chuck lug (22.6);
a chip (1) is arranged in the rectangular inner cavity (22.1) of the chuck (22), one end of the chip (1) is propped against the sliding block (21.2), the other end of the chip (1) is propped against the baffle (22.5), and the spring (3) is in a compressed state;
the emulsion droplet generation unit (13) comprises 4 liquid reservoirs arranged on the top surface of the chip substrate (11) and network through grooves (130) formed in the bottom surface of the chip substrate (11) and communicated with the liquid reservoirs, the 4 liquid reservoirs are distributed on two sides of the chip substrate (11) in two rows, and the 4 liquid reservoirs are an oil phase liquid reservoir (131), a sample liquid reservoir (132), a first collecting liquid reservoir (133) and a second collecting liquid reservoir (134) respectively; the network through tank (130) comprises an oil phase droplet generation tank (135 a) communicated with the oil phase liquid storage tank (131) and a sample droplet generation tank (136 a) communicated with the sample liquid storage tank (132), the oil phase droplet generation tank (135 a) and the sample droplet generation tank (136 a) are intersected in an emulsion generation tank (137 a), the outlet of the emulsion generation tank (137 a) is divided into two paths, one path is communicated with the first collecting liquid storage tank (133), and the other path is communicated with the second collecting liquid storage tank (134); the chip substrate (11) and the chip bottom plate (12) are attached, and then the network through groove (130) forms a sealing pipeline for fluid flow;
an oil phase sample injection micropore (131 a) is formed in the bottom of the oil phase liquid storage pond (131), a sample injection micropore (132 a) is formed in the bottom of the sample liquid storage pond (132), a first collecting micropore (133 a) is formed in the bottom of the first collecting liquid storage pond (133), and a second collecting micropore (134 a) is formed in the bottom of the second collecting liquid storage pond (134); the oil phase droplet generation tank (135 a) is communicated with the oil phase sampling micropore (131 a) through the oil phase liquid inlet tank (135), the sample droplet generation tank (136 a) is communicated with the sample sampling micropore (132 a) through the sample liquid inlet tank (136), and the emulsion generation tank (137 a) is respectively communicated with the first collecting micropore (133 a) and the second collecting micropore (134 a) through the emulsion buffer tank (137); the depth of each through groove in the network through groove (130) is the same and is 10-40 um, the width of the oil phase droplet generation groove (135 a) is 0.2-0.3 times of the oil phase liquid inlet groove (135), the width of the sample droplet generation groove (136 a) is 0.2-0.3 times of the sample liquid inlet groove (136), and the width of the emulsion generation groove (137 a) is 0.2-0.3 times of the emulsion buffer groove (137);
the two oil phase droplet generation tanks (135 a) are arranged, and the two oil phase droplet generation tanks (135 a) are respectively communicated with the oil phase sampling micropores (131 a) through the oil phase liquid inlet tanks (135);
the oil phase sample injection micropore (131 a) and the sample injection micropore (132 a) are respectively provided with a filter column group (138);
4 groups of emulsion droplet generation units (13) are arranged on the chip substrate (11); the 4 groups of emulsion droplet generation units (13) are sequentially and uniformly distributed along the length direction of the chip substrate (11);
the bottom surface of the chip substrate (11) is provided with a recess (11 a), and the top surface of the chip substrate (12) is provided with a protrusion (12 a) corresponding to the recess (11 a).
2. The microdroplet digital PCR chip module as in claim 1, wherein: the chip also comprises a cover plate (4) arranged on the chip (1), wherein the cover plate (4) is provided with a vent hole (41) for injecting gas into the oil phase liquid storage pool (131) and the sample liquid storage pool (132) in the chip (1).
3. The microdroplet digital PCR chip module as claimed in claim 2, wherein: the cover plate (4) is provided with a positioning hole (42), the chuck (22) is provided with a positioning column (22.7), and the positioning column (22.7) penetrates into the positioning hole (42).
4. The microdroplet digital PCR chip module as in claim 1, wherein: the inner sides of the first side (22.2) and the second side (22.3) of the chuck (22) are provided with mounting grooves (22.8) matched with the chip (1), and an arc-shaped positioning groove (22.9) matched with the liquid storage tank is formed in the transverse plate (22.4) of the chuck (22).
5. The microdroplet digital PCR chip module as in claim 1, wherein: the chuck (22) is fixed with the bottom plate (21) through a screw (5).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710106548.8A CN106834115B (en) | 2017-02-27 | 2017-02-27 | Microdroplet digital PCR chip module |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710106548.8A CN106834115B (en) | 2017-02-27 | 2017-02-27 | Microdroplet digital PCR chip module |
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| Publication Number | Publication Date |
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| CN106834115A CN106834115A (en) | 2017-06-13 |
| CN106834115B true CN106834115B (en) | 2023-10-24 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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