CN113980796B - Micro-droplet bearing chip, application thereof, nucleic acid detection module and method - Google Patents
Micro-droplet bearing chip, application thereof, nucleic acid detection module and method Download PDFInfo
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- 238000001514 detection method Methods 0.000 title claims abstract description 65
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 51
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 51
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 51
- 238000000034 method Methods 0.000 title abstract description 6
- 238000007789 sealing Methods 0.000 claims abstract description 11
- 230000005284 excitation Effects 0.000 claims description 25
- 229920003023 plastic Polymers 0.000 claims description 21
- 239000004033 plastic Substances 0.000 claims description 21
- 239000011521 glass Substances 0.000 claims description 19
- 230000003321 amplification Effects 0.000 claims description 13
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 13
- 239000000463 material Substances 0.000 claims description 10
- 239000002184 metal Substances 0.000 claims description 3
- 230000001678 irradiating effect Effects 0.000 claims description 2
- 238000007599 discharging Methods 0.000 abstract description 5
- 238000007847 digital PCR Methods 0.000 description 17
- 239000000758 substrate Substances 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 7
- 238000006243 chemical reaction Methods 0.000 description 6
- 239000007788 liquid Substances 0.000 description 4
- 238000012408 PCR amplification Methods 0.000 description 2
- 238000004891 communication Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 238000000018 DNA microarray Methods 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000006957 competitive inhibition Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011304 droplet digital PCR Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 230000004907 flux Effects 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 238000011901 isothermal amplification Methods 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
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- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
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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
- B01L3/502723—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 characterised by venting arrangements
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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
- B01L3/502769—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 characterised by multiphase flow arrangements
- B01L3/502784—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 characterised by multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
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Abstract
The invention provides a micro-droplet carrying chip, application thereof, a nucleic acid detection module and a method, wherein the micro-droplet carrying chip comprises a chip cover and an oil storage chip which are mutually buckled, a thin layer cavity is formed between the chip cover and the oil storage chip and is used for carrying sample droplets, an oil layer is injected into the thin layer cavity and is used for sealing the sample droplets, and the chip cover is matched with the oil storage chip and is used for detecting nucleic acid of a single sample; the microdroplet-carrying chip also includes a chip carrier having at least one chip site. The chip cover and the oil storage chip provided by the invention can exist independently of the chip frame, can be independently used as a droplet carrying unit for detecting nucleic acid of a single detection sample, and can be used for discharging bubbles by matching with a thin layer cavity formed by the chip cover, so that the generation of droplets is met, the signal detection requirement is met, and the multi-channel nucleic acid detection can be performed on a group sample.
Description
Technical Field
The invention belongs to the technical field of nucleic acid detection, and relates to a micro-droplet carrying chip, application thereof, a nucleic acid detection module and a method.
Background
Currently, digital PCR (dPCR) technology is rapidly evolving as a powerful competitor to fluorescent quantification technology. Various schemes have been proposed to address the problems of droplet loading, droplet amplification, and signal detection in digital PCR. The micro-fluidic chip based on micro-nano processing is an important solution, and comprises a micro-channel droplet generation chip and a micro-pore array chip. This also divides the dPCR platform into two broad categories, one of which is microdroplet digital PCR based on microdroplet technology; and secondly, microwell digital PCR based on microwell technology.
Microdroplet digital PCR is a breakthrough nucleic acid quantitative detection technology developed rapidly in recent years, and a large reaction system is subjected to microdroplet treatment before traditional PCR amplification, and the reaction system is divided into thousands of water-in-oil microdroplets, namely thousands of independent PCR reaction systems. In this process, the sample is diluted to a single molecular level using a droplet forming device and distributed equally among the tens of thousands of reaction systems, each droplet containing no or at least one nucleic acid target molecule to be detected. After amplification, each droplet is detected one by droplet digital PCR, and the original concentration of the sample can be calculated through a Poisson probability distribution formula. Compared with the traditional PCR, the micro-drop digital PCR separates the samples in different reaction systems, effectively avoids the mutual hybridization between different primers or products and the competitive inhibition between the same products in the reaction process, can realize the simultaneous amplification of different templates, and can also obtain higher amplification efficiency. Furthermore, due to the reactive nature of microdroplet digital PCR, the amount of sample required is very small, and the probability of single DNA templates being present in a single microdroplet system without detection is very low, a tremendous improvement over the second generation qPCR. The microdroplet digital PCR has various irreplaceable advantages in the detection field, is particularly suitable for detecting and quantifying trace DNA, is flexible to use, can adjust flux and sensitivity according to experimental requirements, and provides more reliable results for nucleic acid detection. The formation of droplets is a key element in the implementation of the droplet-type digital PCR technology.
CN113278494a digital PCR droplet generation chip comprising, a chip body comprising a sample supply reservoir, at least two independent droplet generation units and a droplet collection reservoir; a droplet generation unit in communication with the sample supply reservoir and the droplet collection reservoir, respectively; the droplet generation unit comprises a micro-channel and a droplet generation part which are communicated in sequence; the cross-sectional area from the inlet end to the outlet end of the droplet generation part is gradually increased, a droplet overflow area is surrounded at the intersection of the outlet end of the micro-channel and the inlet end of the droplet generation part, and the areas of the droplet overflow areas in at least two droplet generation units are different.
CN106834115a discloses a microdroplet type digital PCR chip module, which comprises a chip comprising a chip substrate, a chip base plate, and several groups of emulsion microdroplet generating units disposed 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.
CN205501281U discloses a special biochip of microdroplet digital PCR, including substrate, micropore array piece and cover plate, the substrate openly is equipped with the recess that holds micropore array piece, the cover plate with the front cooperation of substrate will the recess surrounds and forms sealed cavity, the recess bottom surface is equipped with the protruding foot of fixed stay micropore array piece, micropore on the micropore array piece is the through-hole.
From the prior art disclosed at present, there are artifacts in both microdroplet digital PCR and microwell digital PCR, which may ultimately lead to poor and non-uniform stability of microdroplet generation. Meanwhile, the micro-fluidic chip is used as a consumable, and the cost and the batch-to-batch difference limit the application of the digital PCR technology.
Disclosure of Invention
Aiming at the defects existing in the prior art, the invention aims to provide a micro-droplet carrying chip, application thereof, a nucleic acid detection module and a method, wherein the chip cover and the oil storage chip provided by the invention can exist independently of a chip frame, can be independently used as a droplet carrying unit for detecting nucleic acid of a single detection sample, and can be matched with a thin layer cavity formed by the chip cover for discharging bubbles so as to meet the generation of droplets, and simultaneously meet the signal detection requirement, and can also be used for detecting multi-channel nucleic acid of a group sample.
To achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, the invention provides a micro-droplet carrying chip, which comprises a chip cover and an oil storage chip which are mutually buckled, wherein a thin layer cavity is formed between the chip cover and the oil storage chip and is used for carrying sample droplets, an oil layer is injected into the thin layer cavity and is used for sealing the sample droplets, and the chip cover is matched with the oil storage chip and is used for detecting nucleic acid of a single sample;
the micro-droplet bearing chip further comprises a chip frame with at least one chip position, the chip cover and the oil storage chip are buckled and then placed on the chip position, and multichannel nucleic acid detection is carried out on different samples in each chip position.
The chip cover and the oil storage chip provided by the invention can exist independently of the chip frame, can be independently used as a droplet carrying unit for detecting nucleic acid of a single detection sample, and can be used for discharging bubbles by matching with a thin layer cavity formed by the chip cover, so that the generation of droplets is met, the signal detection requirement is met, and the multi-channel nucleic acid detection can be performed on a group sample.
As a preferable technical scheme of the invention, the oil storage chip comprises a side wall and a bottom plate for sealing one end of the side wall.
Preferably, an annular boss is arranged at the outer edge of one end of the side wall, which is far away from the bottom plate.
Preferably, the inner surface of the side wall is provided with an annular step surface, and the annular step surface is clung to the bottom plate.
Preferably, the annular boss, the side wall and the annular step surface are combined to form an annular structure with a Z-shaped section.
As a preferred technical solution of the present invention, the chip cover includes an outer wall and an end cap sealing one end of the outer wall.
Preferably, an annular protruding edge is arranged on the outer wall of the chip cover, which is far away from the outer edge of one end of the end cover, and the annular protruding edge of the chip cover is arranged on the annular boss of the oil storage chip.
As a preferable technical scheme of the invention, the surface of one side of the end cover, which is close to the bottom plate of the oil storage chip, is a plane or a curved surface, and more preferably a curved surface.
Preferably, the center of the surface of the end cover protrudes towards the bottom plate of the oil storage chip.
Preferably, the outer wall of the chip cover is an inverted conical inclined plane, and a reducing annular cavity is formed between the outer wall of the chip cover and the inner surface of the side wall of the oil storage chip.
As a preferable technical scheme of the invention, the chip rack is made of metal or plastic.
Preferably, the material of the oil storage chip comprises glass or light-proof plastic.
Preferably, the material of the chip cover comprises glass or light-proof plastic.
As a preferable technical scheme of the invention, the bottom plate and the side wall of the oil storage chip are made of opaque plastics, the end cover of the chip cover is made of glass, and the outer wall of the chip cover is made of opaque plastics, so that the signal detection mode of upper detection is met.
It should be noted that, the "upper detection" in the present invention means that the excitation light emitter and the camera are both located above the micro-droplet carrying chip, the excitation light emitted by the excitation light emitter passes through the transparent glass end cover from top to bottom and irradiates onto the sample droplet, the fluorescent group in the excited droplet generates emission light under the irradiation of the excitation light, and the outer walls of the oil storage chip and the chip cover are made of opaque plastic materials, so that the emission light emitted by the fluorescent group can only enter the camera through reflection, and the emission light collected by the camera carries out droplet signal detection.
As a preferable technical scheme of the invention, the bottom plate of the oil storage chip is made of glass, the side wall of the oil storage chip is made of opaque plastic, and the end cover and the outer wall of the chip cover are made of opaque glass, so as to meet the signal detection mode of lower detection.
Similarly, the "lower detection" in the present invention means that the excitation light emitter and the camera are both located below the micro-droplet carrying chip, the excitation light emitted by the excitation light emitter irradiates the sample droplet from bottom to top through the transparent glass bottom plate, and the fluorescent group in the excited droplet generates emission light under the irradiation of the excitation light.
In a second aspect, the present invention provides a nucleic acid detection module, where the nucleic acid detection module includes the micro-droplet carrying chip of the first aspect, and an excitation light emitter and a camera are disposed on one side of the micro-droplet carrying chip.
In a third aspect, the present invention provides a nucleic acid detection method of the nucleic acid detection module of the first aspect, the nucleic acid detection method comprising:
and placing the nucleic acid detection module in amplification equipment, performing nucleic acid amplification on the sample microdroplet, irradiating excitation light emitted by an excitation light emitter onto the amplified sample microdroplet after amplification is completed, exciting fluorescent groups in the sample microdroplet to emit emission light, and collecting the emission light through a camera.
In a fourth aspect, the present invention provides a use of the micro-droplet carrier chip according to the first aspect, where the micro-droplet carrier chip is used for detecting nucleic acids.
It should be noted that the application fields of the micro-droplet carrier chip provided by the invention include, but are not limited to, nucleic acid detection based on PCR and isothermal amplification.
Compared with the prior art, the invention has the beneficial effects that:
the chip cover and the oil storage chip provided by the invention can exist independently of the chip frame, can be independently used as a droplet carrying unit for detecting nucleic acid of a single detection sample, and can be used for discharging bubbles by matching with a thin layer cavity formed by the chip cover, so that the generation of droplets is met, the signal detection requirement is met, and the multi-channel nucleic acid detection can be performed on a group sample.
Drawings
FIG. 1 is a cross-sectional view of a chip cover and an oil reservoir chip provided in one embodiment of the invention;
FIG. 2 is a perspective view of a chip cover and an oil storage chip according to an embodiment of the present invention;
FIG. 3 is a schematic diagram illustrating an assembly of a micro-droplet carrier chip according to an embodiment of the present invention;
FIG. 4 is a schematic diagram showing the structure of a nucleic acid detecting module according to an embodiment of the present invention;
wherein, 1-the oil storage chip; 2-a chip cover; 3-chip rack; 4-end caps; 5-an outer wall; 6-annular convex edges; 7-a bottom plate; 8-side walls; 9-an annular boss; 10-an annular step surface; 11-sample droplets; 12-oil layer; 13-activating the light emitter; 14-camera.
Detailed Description
It is to be understood that in the description of the present invention, the terms "center", "longitudinal", "lateral", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc. indicate orientations or positional relationships based on the drawings, are merely for convenience in describing the present invention and simplifying the description, and do not indicate or imply that the devices or elements referred to must have a specific orientation, be configured and operated in a specific orientation, and thus are not to be construed as limiting the present invention. Furthermore, the terms "first," "second," and the like, are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first", "a second", etc. may explicitly or implicitly include one or more such feature. In the description of the present invention, unless otherwise indicated, the meaning of "a plurality" is two or more.
It should be noted that, in the description of the present invention, unless explicitly specified and limited otherwise, the terms "disposed," "connected," and "connected" are to be construed broadly, and may be, for example, fixedly connected, detachably connected, or integrally connected; can be mechanically or electrically connected; can be directly connected or indirectly connected through an intermediate medium, and can be communication between two elements. The specific meaning of the above terms in the present invention can be understood by those of ordinary skill in the art in a specific case.
The technical scheme of the invention is further described by the following specific embodiments.
In a specific embodiment, the invention provides a micro-droplet carrying chip, as shown in fig. 1 and 2, which comprises a chip cover 2 and an oil storage chip 1 which are buckled with each other, wherein a thin layer cavity is formed between the chip cover 2 and the oil storage chip 1 and is used for carrying sample droplets 11, an oil layer 12 is injected into the thin layer cavity, the oil layer 12 is used for sealing the sample droplets 11, and the chip cover 2 is matched with the oil storage chip 1 and is used for detecting nucleic acid of a single sample.
As shown in fig. 3, the micro-droplet carrying chip further includes a chip frame 3 having at least one chip position, the chip cover 2 and the oil storage chip 1 are buckled and then placed on the chip position, and the multi-channel nucleic acid detection is performed on different samples in each chip position.
The chip cover 2 and the oil storage chip 1 provided by the invention can exist independently of the chip frame 3, can be independently used as a droplet carrying unit for detecting nucleic acid of a single detection sample, and the chip cover 2 and a thin layer cavity formed by the cooperation of the chip cover 2 are used for discharging bubbles so as to meet droplet generation, meet the signal detection requirement, and can also be used for detecting multi-channel nucleic acid of a group sample.
Further, the oil storage chip 1 includes a side wall 8 and a bottom plate 7 sealing one end of the side wall 8.
Further, an annular boss 9 is disposed at an outer edge of one end of the side wall 8 away from the bottom plate 7, and the annular flange 6 of the chip cover 2 is mounted on the annular boss 9 of the oil storage chip 1.
Further, an annular step surface 10 is arranged on the inner surface of the side wall 8, and the annular step surface 10 is closely attached to the bottom plate 7.
Further, the annular boss 9, the side wall 8 and the annular step surface 10 are combined to form an annular structure with a Z-shaped section.
Further, the chip cover 2 includes an outer wall 5 and an end cap 4 sealing one end of the outer wall 5.
Further, an annular protruding edge 6 is arranged at the outer edge of one end, far away from the end cover 4, of the outer wall 5 of the chip cover 2, and the annular protruding edge 6 of the chip cover 2 is arranged on an annular boss 9 of the oil storage chip 1.
Further, a surface of the end cover 4, which is close to the bottom plate 7 of the oil storage chip 1, is a plane or a curved surface, and more preferably a curved surface.
Further, the center of the surface of the end cover 4 protrudes toward the bottom plate 7 of the oil storage chip 1.
Further, the outer wall 5 of the chip cover 2 is an inverted conical inclined surface, and a reducing annular cavity is formed between the outer wall 5 of the chip cover 2 and the inner surface of the side wall 8 of the oil storage chip 1.
Further, the material of the chip frame 3 is metal or plastic.
Further, the material of the oil storage chip 1 comprises glass or light-proof plastic.
Further, the material of the chip cover 2 includes glass or opaque plastic.
Further, the bottom plate 7 and the side wall 8 of the oil storage chip 1 are made of opaque plastics, the end cover 4 of the chip cover 2 is made of glass, and the outer wall 5 of the chip cover 2 is made of opaque plastics, so as to meet the signal detection mode of upper detection.
It should be noted that, in the present invention, "upper detection" means that the excitation light emitter 13 and the camera 14 are located above the micro-droplet carrying chip, the excitation light emitted by the excitation light emitter 13 passes through the transparent glass end cover 4 from top to bottom and irradiates onto the sample droplet 11, and the fluorescent groups in the excited droplet generate emission light under the irradiation of the excitation light, and since the oil storage chip 1 and the outer wall 5 of the chip cover 2 are made of opaque plastic materials, the emission light emitted by the fluorescent groups can only enter the camera 14 through reflection, and the droplet signal detection is performed by the emission light collected by the camera 14.
Further, the bottom plate 7 of the oil storage chip 1 is made of glass, the side wall 8 of the oil storage chip 1 is made of opaque plastic, and the end cover 4 and the outer wall 5 of the chip cover 2 are made of opaque glass, so as to meet the signal detection mode of lower detection.
Similarly, the "lower detection" in the present invention means that the excitation light emitter 13 and the camera 14 are both located below the micro-droplet carrying chip, the excitation light emitted by the excitation light emitter 13 irradiates onto the sample droplet 11 from bottom to top through the transparent glass bottom plate 7, and the fluorescent groups in the excited droplet generate emission light under the irradiation of the excitation light, and since the chip cover 2 and the side wall 8 of the oil storage chip 1 are made of opaque plastic materials, the emission light emitted by the fluorescent groups can only enter the camera 14 through reflection, and the droplet signal detection is performed by the emission light collected by the camera 14.
In another embodiment, the invention provides a nucleic acid detection module, as shown in fig. 4, which comprises the micro-droplet carrying chip provided in the embodiment, and one side of the micro-droplet carrying chip is provided with an excitation light emitter 13 and a camera 14.
In another embodiment, the present invention provides a nucleic acid detection method of a nucleic acid detection module, the nucleic acid detection method comprising: the nucleic acid detection module is placed in amplification equipment, nucleic acid amplification is carried out on the sample droplet 11, after amplification is finished, excitation light emitted by the excitation light emitter 13 irradiates the amplified sample droplet 11, fluorescent groups in the sample droplet 11 are excited to emit emission light, and the emission light is collected by the camera 14.
In another embodiment, the invention provides a use of the micro-droplet carrying chip for detecting nucleic acid.
The applicant declares that the above is only a specific embodiment of the present invention, but the scope of the present invention is not limited thereto, and it should be apparent to those skilled in the art that any changes or substitutions that are easily conceivable within the technical scope of the present invention disclosed by the present invention fall within the scope of the present invention and the disclosure.
Claims (13)
1. The micro-droplet bearing chip is characterized by comprising a chip cover and an oil storage chip which are mutually buckled, wherein a thin-layer cavity is formed between the chip cover and the oil storage chip and is used for bearing sample droplets, an oil layer is injected into the thin-layer cavity and is used for sealing the sample droplets, and the chip cover is matched with the oil storage chip and is used for detecting nucleic acid of a single sample;
the oil storage chip comprises a side wall and a bottom plate for sealing one end of the side wall, an annular boss is arranged at the outer edge of one end, far away from the bottom plate, of the side wall, an annular step surface is arranged on the inner surface of the side wall, the annular step surface is tightly attached to the bottom plate, and the annular boss, the side wall and the annular step surface are combined to form an annular structure with a Z-shaped section;
the chip cover comprises an outer wall and an end cover for sealing one end of the outer wall, an annular convex edge is arranged at the outer edge of one end, far away from the end cover, of the outer wall of the chip cover, and the annular convex edge of the chip cover is arranged on an annular boss of the oil storage chip;
the micro-droplet bearing chip further comprises a chip frame with at least one chip position, the chip cover and the oil storage chip are buckled and then placed on the chip position, and multichannel nucleic acid detection is carried out on different samples in each chip position.
2. The micro-droplet carrier chip of claim 1, wherein a surface of the end cap adjacent to the bottom plate of the oil storage chip is a plane or a curved surface.
3. The micro-droplet carrier chip of claim 1, wherein a surface of the end cap adjacent to the bottom plate of the oil storage chip is curved.
4. The micro-droplet carrier chip of claim 1, wherein the center of the end cap surface is raised toward the reservoir chip floor.
5. The micro-droplet carrying chip according to claim 1, wherein the outer wall of the chip cover is an inverted conical inclined surface, and a reducing annular cavity is formed between the outer wall of the chip cover and the inner surface of the side wall of the oil storage chip.
6. The micro-droplet carrier chip of claim 1, wherein the material of the chip carrier is metal or plastic.
7. The micro-droplet carrier chip of claim 1, wherein the oil reservoir chip comprises glass or light-impermeable plastic.
8. The micro-droplet carrier chip of claim 1, wherein the material of the chip cover comprises glass or light-impermeable plastic.
9. The micro-droplet carrying chip according to claim 1, wherein the bottom plate and the side wall of the oil storage chip are made of opaque plastics, the end cover of the chip cover is made of glass, and the outer wall of the chip cover is made of opaque plastics for satisfying a signal detection mode of upper detection.
10. The micro-droplet carrying chip according to claim 1, wherein the bottom plate of the oil storage chip is made of glass, the side wall of the oil storage chip is made of opaque plastic, and the end cover and the outer wall of the chip cover are made of opaque glass for satisfying a signal detection mode of lower detection.
11. A nucleic acid detection module, characterized in that the nucleic acid detection module comprises the micro-droplet carrying chip according to any one of claims 1 to 10, wherein an excitation light emitter and a camera are arranged on one side of the micro-droplet carrying chip.
12. A nucleic acid detection method of the nucleic acid detection module of claim 11, wherein the nucleic acid detection method comprises:
and placing the nucleic acid detection module in amplification equipment, performing nucleic acid amplification on the sample microdroplet, irradiating excitation light emitted by an excitation light emitter onto the amplified sample microdroplet after amplification is completed, exciting fluorescent groups in the sample microdroplet to emit emission light, and collecting the emission light through a camera.
13. Use of a micro-droplet carrier chip according to any of claims 1-10 for nucleic acid detection.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111243192.5A CN113980796B (en) | 2021-10-25 | 2021-10-25 | Micro-droplet bearing chip, application thereof, nucleic acid detection module and method |
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