CN111304072B - Oil seal device for digital PCR chip - Google Patents
Oil seal device for digital PCR chip Download PDFInfo
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- CN111304072B CN111304072B CN202010128380.2A CN202010128380A CN111304072B CN 111304072 B CN111304072 B CN 111304072B CN 202010128380 A CN202010128380 A CN 202010128380A CN 111304072 B CN111304072 B CN 111304072B
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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
- C12Q1/6851—Quantitative amplification
Abstract
The invention provides an oil seal device for a digital PCR chip, which comprises: the substrate is made of aluminum, a groove is formed in the middle of the substrate and is used as a container for placing a reaction chip, an upper cover is arranged on the groove, the upper cover is slid relative to the substrate to seal the chip, and paraffin oil with the thickness of 0.3-0.5mm is used as an oil seal structure at the position corresponding to the chip by the upper cover; the two opposite sides of the inner side of the groove are provided with sliding grooves, the upper cover can horizontally slide in the sliding grooves, and the upper side surface of the side wall of the sliding groove is provided with an oil cover. The fully-closed and semi-automatic operation reduces the manual operation process, so that errors caused by manual operation can be reduced to the greatest extent; good heat conducting medium is selected to fully ensure the heat conductivity of the device as a reaction chip container, and the stability of reaction conditions is fully ensured; the upper cover surface is made of materials and is of a structure, so that the effectiveness of later-stage result detection can be fully guaranteed.
Description
Technical Field
The invention relates to the technical field of oil sealing of nucleic acid amplification instruments, in particular to an oil sealing device for a digital PCR chip.
Background
The use of DNA molecular amplification technology has prompted the development of molecular biology techniques, and accurate quantification of DNA copy number is one of the important applications of modern molecular biology and medicine. In 1997, james F.brown, jonathan E.silver and Olga V.kalina constructed a prototype of the digital PCR (digital polymerase chain reaction, dPCR) technique and obtained U.S. Pat. No. US 6143496A. Different from the traditional PCR technology, the digital PCR sufficiently dilutes the nucleic acid samples, so that the number of sample templates of each reaction chamber is less than or equal to 1, and the fluorescent signals of each reaction chamber are subjected to statistical analysis after the PCR amplification is finished, so that a series of problems in the relative quantitative process of real-time fluorescent quantitative PCR (real time quantitative PCR, qPCR) by using a reference standard sample and a standard curve are solved, and absolute quantification of single-molecule DNA is realized. The technology has high sensitivity, strong specificity, high detection flux and accurate quantification, and has been widely applied to the aspects of clinical diagnosis, transgene component quantification, single cell gene expression, environmental microorganism detection, next generation sequencing and the like.
The digital PCR (digital PCR-dPCR) technology is a new nucleic acid detection and quantification method, and unlike the traditional quantitative PCR (qPCR) technology, the digital PCR adopts an absolute quantification mode, and directly detects the copy number of a target sequence without depending on a standard curve and a reference sample. Because the detection mode has more excellent sensitivity, specificity and accuracy than the traditional qPCR, dPCR is rapidly and widely applied, the technology has been widely accepted in terms of very trace nucleic acid sample detection, rare mutation detection under a complex background and small difference identification of expression quantity, and has a wide application prospect in the aspects of gene expression research, microRNA research, genome copy number identification, cancer marker rare mutation detection, pathogenic microorganism identification, transgene component identification, accurate quantification of NGS sequencing library, result verification and the like, and has been paid more attention. Digital PCR techniques are currently classified into two types, chip-type and droplet-type, in which chip-type digital PCR devices and techniques employ chips as reaction vessels for nucleic acid amplification, which require that the nucleic acid amplification reaction must be performed in a fully enclosed environment due to the nature of digital PCR techniques and real-time fluorescent quantitative PCR techniques themselves.
At present, a chip carrying device of a chip type digital PCR (digital PCR chip carrying device of Thermo Scientific company) adopts aluminum as a substrate, plastic and glass covers are used as sealing covers on the upper surface, and a mode of scraping mixed liquid of a sample and a reaction reagent into a micro-reaction tank by using a special scraper is needed for chip sample adding. And (3) filling mineral oil for sealing after sample addition, and then placing an amplification instrument for nucleic acid amplification reaction after the upper surface sealing cover is attached to the aluminum substrate by using strong glue.
The main defects of the operation are that the steps of the operation process are more, the process is complex, certain technical requirements are provided for operators, the possibility of human error is high, the operation time is long, and the mixed liquid in the reaction tank of the micro system is possibly volatilized, so that the result is inaccurate.
Disclosure of Invention
The invention aims to provide an oil sealing device for a digital PCR chip, so as to solve the technical problems.
In order to achieve the above object, the present invention provides an oil seal device for a digital PCR chip, comprising: the substrate is made of aluminum, a groove is formed in the middle of the substrate and is used as a container for placing a reaction chip, an upper cover is arranged on the groove, the upper cover is slid relative to the substrate to seal the chip, and paraffin oil with the thickness of 0.3-0.5mm is used as an oil seal structure at the position corresponding to the chip by the upper cover;
the upper cover can horizontally slide in the chute, and the upper side surface of the side wall of the chute is provided with an oil cover;
one side of recess is provided with an infrared inductor, detects the height h of the oil front cover in the recess through the emission infrared ray, and the paraffin is injected through the oiling machine to can confirm the discharge capacity Q of paraffin in real time, the oiling machine can confirm the volume V of waiting to inject the paraffin through the discharge capacity Q of injecting the paraffin and the height h of oil front cover.
Further, after the upper cover is in place, the oiling machine injects paraffin oil displacement Q1 into the groove, the infrared sensor detects the oil surface height h1 of the oil cover at the moment, and the infrared sensor acquires the oil surface height h1 and then transmits the oil surface height h1 to the controller to be compared with preset displacement height information; and a preset first oil level height h10 is set in the controller, and the oil level height h1 measured in real time is compared with the preset first oil level height h10 so as to acquire oil seal information of the upper cover.
Further, comparing the oil level height h1 measured in real time with a preset first oil level height h10, if the difference (h 1-h 10) between the first oil level height h1 and the preset first oil level height h10 is greater than 0.5xh10, the upper cover is not completely sealed, paraffin oil leaks downwards through the upper cover, at the moment, the upper cover is required to be readjusted and opened, the chip is required to be sealed again by the upper cover, and paraffin oil is injected through the oiling machine.
Further, if the difference between the first oil level h1 and the preset first oil level h10 is 0.2xh10 < (h 1-h 10) < 0.5xh10, at this time, 0.5xq1 is injected into the groove through the oil injector to measure the sealing condition of the upper cover, and after paraffin oil of 0.5xq1 is injected, the infrared sensor measures the oil level h11.
Further, if the oil level h11 measured in real time is smaller than the preset first oil level h10, the upper cover is not completely sealed, paraffin oil leaks downwards through the upper cover, at the moment, the upper cover is required to be readjusted and opened and the chip is required to be sealed again by the upper cover, and then the paraffin oil is injected through the oiling machine; if the oil level height h11 measured in real time is greater than the preset first oil level height h10 and less than 1.3x h10, the upper cover is completely sealed, and after the paraffin oil is injected for the first time, the paraffin oil fills the leak, and at the moment, the oil seal is in place.
Further, if the difference (h 1-h 10) between the first oil level h1 and the preset first oil level h10 is less than 0.2xh10, at this time, the controller acquires the first oil level height curve, and if there is no trough at the edge of the first oil level height curve, there is no gap between the upper cover and the groove, or after the first oil injection, the gap filling between the upper cover and the side wall of the groove is completed, and paraffin oil is continuously injected through the oiling machine, so as to complete the oil sealing process.
Further, according to the height of the first oil level height curve, the height value of the paraffin oil at the height of the curve is obtained, if a trough exists at the edge of the first oil level height curve, a gap exists between the upper cover and the groove, and the upper cover needs to be sealed again.
Further, the oiling machine continuously injects paraffin oil with the discharge capacity Q2 into the groove, the infrared sensor acquires the oil surface height h2 of the oil cover in real time, compares the oil surface height h2 with preset discharge capacity height information, sets the second oil surface height h20, and compares the oil surface height h2 measured in real time with the preset second oil surface height h 20.
Further, if the difference (h 2-h 20) between the oil level height measured in real time and the preset second oil level height h20 is smaller than 0.1xh2, acquiring a second oil level height curve through an infrared sensor during the second injection displacement Q2, acquiring the height value of paraffin oil at the height of the curve according to the height of the second oil level height curve, and if a trough exists at the edge of the curve, forming a gap between an upper cover and a groove at the position, and re-covering the upper cover; if no trough exists at the edge of the curve, no gap exists between the upper cover and the groove, and the oil seal process is completed.
Further, if the difference (h 2-h 20) between the oil level height measured in real time and the preset second oil level height h20 is greater than 0.1xh2, the oil seal does not meet the requirement, at this time, 0.5x Q2 is injected into the groove through the oiling machine, the infrared sensor measures the oil level height h12, and if at this time, the oil level height h12 measured in real time is smaller than the preset second oil level height h20, the upper cover is required to be readjusted and opened, the chip is required to be sealed again by the upper cover, and then paraffin oil is injected through the oiling machine; if the oil level height h12 measured in real time is larger than the preset second oil level height h20, and the trough does not exist at the edge of the re-measured third oil level curve, the oil seal is carried out according to the paraffin oil thickness at the moment.
Compared with the prior art, the oil sealing device for the PCR chip has the technical effects that the process of manual operation is reduced due to the full-sealing and semi-automatic operation, so that errors caused by the manual operation can be reduced to the greatest extent; good heat conducting medium is selected to fully ensure the heat conductivity of the device as a reaction chip container, and the stability of reaction conditions is fully ensured; the upper cover surface is made of materials and is of a structure, so that the effectiveness of later-stage result detection can be fully guaranteed.
In particular, the invention judges the edge valley of the oil seal curve of the oil seal and the height information of the oil seal to obtain the real-time upper cover sealing condition and make corresponding reaction so as to achieve perfect paraffin oil sealing.
Particularly, after the upper cover is in place, the oiling machine injects paraffin oil into the groove to discharge Q1, the infrared sensor detects the oil surface height h1 of the oil cover at the moment, and the infrared sensor acquires the oil surface height h1 and then transmits the oil surface height h1 into the controller to be compared with preset discharge height information; a preset first oil level height h10 is set in the controller, and the oil level height h1 measured in real time is compared with the preset first oil level height h10 to obtain oil seal information of the upper cover; comparing the oil level height h1 measured in real time with a preset first oil level height h10, if the difference (h 1-h 10) between the first oil level height h1 and the preset first oil level height h10 is greater than 0.5xh10, the upper cover is not completely sealed, paraffin oil leaks downwards through the upper cover, at the moment, the upper cover is required to be readjusted and opened, the chip is required to be sealed again by the upper cover, and paraffin oil is injected through the oiling machine; if the difference between the first oil level h1 and the preset first oil level h10 is 0.2xh10 < (h 1-h 10) < 0.5xh10, at this time, 0.5xQ1 is injected into the groove through the oiling machine to measure the sealing condition of the upper cover, and after 0.5xQ1 of paraffin oil is injected, the infrared sensor measures the oil level h11.
Drawings
Various other advantages and benefits will become apparent to those of ordinary skill in the art upon reading the following detailed description of the preferred embodiments. The drawings are only for purposes of illustrating the preferred embodiments and are not to be construed as limiting the invention. Also, like reference numerals are used to designate like parts throughout the figures. In the drawings:
FIG. 1 is a schematic diagram of an oil seal device for a digital PCR chip according to an embodiment of the present invention;
FIG. 2 is a schematic diagram of a cover plate structure according to an embodiment of the invention;
fig. 3 is a schematic structural diagram of an oil cover according to an embodiment of the invention.
Detailed Description
Preferred embodiments of the invention are described below with reference to the accompanying drawings. It should be understood by those skilled in the art that these embodiments are merely for explaining the technical principles of the present invention, and are not intended to limit the scope of the present invention.
It should be noted that, in the description of the present invention, terms such as "upper," "lower," "left," "right," "inner," "outer," and the like indicate directional or positional relationships based on the directional or positional relationships shown in the drawings, which are merely for convenience of description, and do not indicate or imply that the apparatus or elements must have a specific orientation, be constructed and operated in a specific orientation, and thus should not be construed as limiting the invention.
Furthermore, it should be noted that, in the description of the present invention, unless explicitly specified and limited otherwise, the terms "mounted," "connected," and "connected" are to be construed broadly, and may be either fixedly connected, detachably connected, or integrally connected, for example; 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 invention will be understood by those skilled in the art according to the specific circumstances.
Referring to fig. 1, which is a schematic diagram of an oil seal device for a digital PCR chip according to an embodiment of the present invention, the oil seal device of the present embodiment includes a base 1 made of aluminum material, a groove 2 is provided in a middle portion of the base 1, the groove is used as a container for placing a reaction chip, an upper cover 3 is provided on the groove 2, in the present embodiment, the upper cover 3 is made of organic glass (PMMA), and paraffin oil with a thickness of 0.3-0.5mm is used as an oil seal structure at a position corresponding to the chip on the upper cover.
Specifically, the upper cover and the substrate of the embodiment are connected by adopting a scribing type structure, the upper cover can relatively slide with the substrate to realize the sealing of the chip, and when the upper cover is buckled on the upper side of the chip, the upper cover is in oil seal through paraffin oil. The substrate of this example was made of aluminum material and the grooves were cut out by digging out the center of the aluminum block to a size of 10x 10 mm.
Specifically, in the process of reacting to the digital PCR chip, the operation procedure of this embodiment is: preparing an oil sealing device and a PCR reaction chip, placing the PCR reaction chip into a groove, injecting a nucleic acid amplification reaction mixed solution into the surface of the reaction chip by using a liquid-transferring gun, slowly pushing the upper cover surface into the groove for sealing, injecting mineral oil into a spare gap, and performing amplification reaction on the machine.
Referring to fig. 2, which is a schematic diagram of an upper cover sliding structure according to an embodiment of the present invention, in conjunction with fig. 1, the upper cover of the present embodiment is a push-pull structure, sliding grooves 23 are disposed on two opposite sides of an inner side of a groove 22, the upper cover 3 can move horizontally in the sliding grooves 23, and after the upper cover 3 slides into the sliding grooves 23, an oil seal is formed on an upper cover by paraffin oil, wherein an oil seal surface 21 is formed on an upper side surface of a side wall of the sliding groove 23, the paraffin oil seal is formed on the oil seal surface, and real-time thickness of the paraffin oil can be embodied on the oil seal surface. Correspondingly, the relative connection structure of the chute and the upper cover can be further provided with a positioning structure and a locking structure, so that the sliding between the chute and the upper cover can be positioned and locked, and as can be understood by those skilled in the art, the positioning structure can be provided with a locking column on the side wall of the groove, and the locking structure can be provided with a lock catch arranged on the upper cover, so that the upper cover is locked, which is a conventional technology and will not be repeated.
Referring to fig. 3, a schematic diagram of an oil cover according to an embodiment of the present invention is shown, where in the process of injecting paraffin oil, the thickness of the injected paraffin oil is controlled in real time to determine the oil seal condition of the upper cover, and also determine the sealing condition of the upper cover. Referring to fig. 1, an infrared sensor 4 is disposed on one side of the groove in the present embodiment, and detects the height h of the oil cover 21 in the groove by emitting infrared rays, so that paraffin oil is injected through the oiling machine, and the displacement Q of the paraffin oil can be determined in real time, and the oiling machine can determine the amount V of the paraffin oil to be injected through the displacement Q of the injected paraffin oil and the height h of the oil cover.
Specifically, in the embodiment of the present invention, after the upper cover is in place, the oil filling machine fills paraffin oil displacement Q1 into the groove, the infrared sensor detects the oil level height h1 of the oil cover 21 at this time, and the infrared sensor acquires the oil level height h1 and then transmits the oil level height h1 to the controller to compare with preset displacement height information. In this embodiment, a preset first oil level h10 is set in the controller, the oil level h1 measured in real time is compared with the preset first oil level h10,
if the difference (h 1-h 10) between the first oil level h1 and the preset first oil level h10 is greater than 0.5x h10, the upper cover is not completely sealed, paraffin oil leaks downwards through the upper cover, at this time, the upper cover needs to be readjusted and opened again, the upper cover is used for sealing the chip again, and then the paraffin oil is injected through the oiling machine.
If the difference between the first oil level h1 and the preset first oil level h10 is 0.2xh10 < (h 1-h 10) < 0.5xh10, the upper cover may not be completely sealed, and may leak through the upper cover, at this time, 0.5xq1 is injected into the groove through the oiling machine to measure the sealing condition of the upper cover, after 0.5xq1 of paraffin oil is injected, the infrared sensor measures the oil level h11, and if at this time, the oil level h11 measured in real time is smaller than the preset first oil level h10, the upper cover fails to be completely sealed, paraffin oil leaks downwards through the upper cover, at this time, the upper cover needs to be readjusted and the upper cover is used again to seal the chip, and then the paraffin oil is injected through the oiling machine. If the oil level height h11 measured in real time is greater than the preset first oil level height h10 and less than 1.3x h10, the upper cover is completely sealed, and after the paraffin oil is injected for the first time, the paraffin oil fills the leak, and at the moment, the oil seal is in place.
If the difference (h 1-h 10) between the first oil level h1 and the preset first oil level h10 is < 0.2xh10, the controller acquires a first oil level height curve 51, where the first oil level height is acquired by an average value of the measured first oil level height curves. In this embodiment, the infrared sensor is disposed at one side of the groove, the first oil level height curve is disposed along the direction shown in fig. 3, and the height value of the paraffin oil at the height of the curve is obtained according to the height of the first oil level height curve, if there is a trough at the edge of the curve, there is a gap between the upper cover and the groove, and the upper cover needs to be capped again; if no trough exists at the edge of the curve, no gap exists between the upper cover and the groove, or gap filling between the upper cover and the side wall of the groove is completed after the first oil injection, paraffin oil is continuously injected through the oil injection machine, and the oil sealing process is completed.
Specifically, the oiling machine continuously injects paraffin oil with the displacement Q2 into the groove, and the infrared sensor acquires the oil surface height h2 of the oil cover 21 in real time and compares the oil surface height h2 with preset displacement height information. In this embodiment, the second oil level h20 is set, the oil level h2 measured in real time is compared with the preset second oil level h20,
if the difference (h 2-h 20) between the oil level height measured in real time and the preset second oil level height h20 is less than 0.1xh2, acquiring a second oil level height curve 52 through an infrared sensor during the second injection displacement Q2, acquiring the height value of paraffin oil at the height of the curve according to the height of the second oil level height curve, and if a trough exists at the edge of the curve, forming a gap between an upper cover and a groove at the position, and re-covering the upper cover; if no trough exists at the edge of the curve, no gap exists between the upper cover and the groove, the oil seal process is completed, and the amplification reaction can be carried out.
If the difference (h 2-h 20) between the oil level height measured in real time and the preset second oil level height h20 is greater than 0.1xh2, the oil seal does not meet the requirement, at the moment, 0.5x Q2 is injected into the groove through the oiling machine, the infrared sensor measures the oil level height h12, and if the oil level height h12 measured in real time is smaller than the preset second oil level height h20 at the moment, the upper cover is required to be readjusted and opened, the chip is sealed by the upper cover again, and then paraffin oil is injected through the oiling machine; if the oil level height h12 measured in real time is greater than the preset second oil level height h20, and the trough does not exist at the edge of the re-measured third oil level curve, the amplification reaction can be performed according to the paraffin oil thickness seal at the moment.
Specifically, the invention judges the edge valley of the oil seal curve of the oil seal and the height information of the oil seal to obtain the real-time upper cover sealing condition and make corresponding reaction so as to achieve perfect paraffin oil sealing.
Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions may be made therein without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (10)
1. An oil seal device for a digital PCR chip, comprising:
the substrate is made of aluminum, a groove is formed in the middle of the substrate and is used as a container for placing a reaction chip, an upper cover is arranged on the groove, the upper cover is slid relative to the substrate to seal the chip, and paraffin oil with the thickness of 0.3-0.5mm is used as an oil seal structure at the position corresponding to the chip by the upper cover;
the upper cover can horizontally slide in the chute, and the upper side surface of the side wall of the chute is provided with an oil cover;
one side of recess is provided with an infrared inductor, detects the height h of the oil front cover in the recess through the emission infrared ray, and the paraffin is injected through the oiling machine to can confirm the discharge capacity Q of paraffin in real time, the oiling machine can confirm the volume V of waiting to inject the paraffin through the discharge capacity Q of injecting the paraffin and the height h of oil front cover.
2. The oil seal device for the digital PCR chip according to claim 1, wherein after the upper cover is in place, the oil injector injects paraffin oil with a displacement of Q1 into the groove, the infrared sensor detects the oil level height h1 of the oil cover at the moment, and the infrared sensor acquires the oil level height h1 and then transmits the oil level height h1 to the controller to be compared with preset displacement height information; and a preset first oil level height h10 is set in the controller, and the oil level height h1 measured in real time is compared with the preset first oil level height h10 so as to acquire oil seal information of the upper cover.
3. The oil seal device for a digital PCR chip according to claim 2, wherein the oil level h1 measured in real time is compared with a preset first oil level h10, and if the difference (h 1-h 10) between the first oil level h1 and the preset first oil level h10 is greater than 0.5x h10, the upper cover is not completely sealed, and paraffin oil leaks downward through the upper cover, and at this time, the upper cover is readjusted to open and the chip is sealed again with the upper cover, and paraffin oil is injected through the oil injector.
4. The oil seal device for digital PCR chip according to claim 2, wherein if the difference between the first oil level h1 and the preset first oil level h10 is 0.2xh10 < (h 1-h 10) < 0.5xh10, the oil level h11 is measured by the infrared sensor after the paraffin oil of 0.5xq1 is injected into the groove by the oil injector to measure the sealing condition of the upper cover.
5. The oil seal device for digital PCR chip as set forth in claim 4, wherein if the oil level h11 measured in real time is less than the preset first oil level h10, the upper cover is not completely sealed, the paraffin oil leaks downward through the upper cover, and at this time, the upper cover is readjusted and the chip is sealed again with the upper cover, and the paraffin oil is injected through the oil injector; if the oil level height h11 measured in real time is greater than the preset first oil level height h10 and less than 1.3x h10, the upper cover is completely sealed, and after the paraffin oil is injected for the first time, the paraffin oil fills the leak, and at the moment, the oil seal is in place.
6. The oil seal device for a digital PCR chip according to claim 2, wherein if the difference (h 1-h 10) between the first oil level h1 and the preset first oil level h10 is < 0.2xh10, the controller obtains a first oil level height curve, and if there is no trough at the edge of the first oil level height curve, there is no gap between the upper cover and the groove, or after the first oil injection, the gap filling between the upper cover and the side wall of the groove is completed, and then the paraffin oil is continuously injected through the oil injection machine to complete the oil seal process.
7. The oil seal device for digital PCR chip as set forth in claim 6, wherein the paraffin oil is obtained according to the height of the first oil level curve, and if there is a trough at the edge of the first oil level curve, there is a gap between the upper cover and the groove, and the upper cover needs to be sealed again.
8. The oil seal device for the digital PCR chip as set forth in claim 6, wherein the oil injection machine continuously injects paraffin oil with a discharge capacity Q2 into the groove, the infrared sensor acquires an oil level height h2 of the oil cover in real time, compares the oil level height h2 with preset discharge capacity height information, sets a second oil level height h20, and compares the oil level height h2 measured in real time with the preset second oil level height h 20.
9. The oil seal device for digital PCR chip according to claim 8, wherein if the difference (h 2-h 20) between the oil level height measured in real time and the preset second oil level height h20 is <0.1xh2, the second oil level height curve is obtained through the infrared sensor during the second injection of the displacement Q2, the height value of the paraffin oil at the height of the curve is obtained according to the height of the second oil level height curve, and if there is a trough at the edge of the curve, there is a gap between the upper cover and the groove, and the upper cover needs to be sealed again; if no trough exists at the edge of the curve, no gap exists between the upper cover and the groove, and the oil seal process is completed.
10. The oil seal device for digital PCR chip according to claim 8, wherein if the difference (h 2-h 20) between the oil level height measured in real time and the preset second oil level height h20 is greater than 0.1xh2, the oil seal is not required, at this time, 0.5x Q2 is injected into the groove through the oil injection machine, the infrared sensor measures the oil level height h12, and if the oil level height h12 measured in real time is less than the preset second oil level height h20, the upper cover is readjusted and the chip is sealed again with the upper cover, and then paraffin oil is injected through the oil injection machine; if the oil level height h12 measured in real time is larger than the preset second oil level height h20, and the trough does not exist at the edge of the re-measured third oil level curve, the oil seal is carried out according to the paraffin oil thickness at the moment.
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