CN110846206A - Full-automatic integrated digital PCR device - Google Patents
Full-automatic integrated digital PCR device Download PDFInfo
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- CN110846206A CN110846206A CN201911241203.9A CN201911241203A CN110846206A CN 110846206 A CN110846206 A CN 110846206A CN 201911241203 A CN201911241203 A CN 201911241203A CN 110846206 A CN110846206 A CN 110846206A
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L7/00—Heating or cooling apparatus; Heat insulating devices
- B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- 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
- B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
- B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates
- B01L3/50851—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates specially adapted for heating or cooling samples
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Abstract
The invention discloses a full-automatic integrated digital PCR device, which comprises a liquid drop generating assembly, a thermal circulation drainage channel, a thermal circulation assembly, a fluorescence detection assembly and a controller, wherein the liquid drop generating assembly is arranged on the thermal circulation drainage channel; the thermal cycle component comprises a liquid drop slide, a mechanical arm, a high-temperature bath kettle and a low-temperature bath kettle; the liquid drops to be detected generated by the liquid drop generating assembly flow into the liquid drop chip through the thermal circulation drainage channel; the mechanical arm is used for circularly switching the liquid drop slide between the high-temperature bath kettle and the low-temperature bath kettle; the fluorescence detection assembly is used for carrying out fluorescence detection on the droplet slide glass after thermal cycling for preset times; the controller is used for controlling the liquid drop generation assembly, the thermal circulation assembly and the fluorescence detection assembly and carrying out quantitative analysis on detection results. According to the invention, the liquid drop to be detected is heated more uniformly, and the finally obtained nucleic acid amplification in the liquid drop to be detected is closer to the ideal condition in experimental design, so that the detection accuracy is higher.
Description
Technical Field
The invention relates to the field of life science, in particular to a full-automatic integrated digital PCR device.
Background
DPCR (digital PCR) is a short name for digital PCR, an absolute quantification technique of nucleic acid molecules, and is the third generation PCR technique. The number of DNA in the starting sample can be directly derived by digital PCR techniques. Dividing a sample into hundreds to tens of thousands of parts, distributing the parts into different reaction units, wherein one or more units contain a target molecule (DNA template), carrying out PCR amplification on the target molecule in each reaction unit respectively, carrying out statistical analysis on fluorescence signals of each reaction unit after the amplification is finished, determining how many DNAs are in the sample by recording the number of DNA reaction units with fluorescence signals, and determining the concentration of the DNAs in an unknown sample by calculating the proportion of the units with fluorescence signals in all the reaction units. At present, the digital PCR can be applied to the detection of pathogens, the detection of cancer cells, gene expression analysis, environmental monitoring, food detection and other aspects, and has wide prospects.
The existing digital PCR device is mostly based on the traditional Tec (semiconductor refrigerator) temperature rise and fall technology, which needs to be matched with peripheral power supply, control circuit, radiator and the like, the whole equipment volume is large, and the Tec temperature rise and fall technology is difficult to uniformly heat a large amount of liquid drops to be tested, therefore, finding a method which can stably control the temperature of each stage of thermal cycle and uniformly heat a large amount of liquid drops to be tested is a problem to be solved urgently by technical personnel in the field.
Disclosure of Invention
The invention aims to provide a full-automatic integrated digital PCR device to solve the problem of inaccurate temperature control of a thermocycling assembly in the prior art.
In order to solve the technical problem, the invention provides a full-automatic integrated digital PCR device, which comprises a liquid drop generating assembly, a thermal cycle drainage channel, a thermal cycle assembly, a fluorescence detection assembly and a controller, wherein the liquid drop generating assembly is connected with the thermal cycle drainage channel;
the thermal cycle component comprises a liquid drop slide, a mechanical arm, a high-temperature bath kettle and a low-temperature bath kettle;
the liquid drops to be detected generated by the liquid drop generating assembly flow into the liquid drop chip through the thermal circulation drainage channel;
the mechanical arm is used for circularly switching the liquid drop slide between the high-temperature bath kettle and the low-temperature bath kettle;
the fluorescence detection assembly is used for carrying out fluorescence detection on the droplet slide glass after thermal cycling for preset times;
the controller is used for controlling the liquid drop generation assembly, the thermal circulation assembly and the fluorescence detection assembly and carrying out quantitative analysis on detection results.
Optionally, in the fully automatic integrated digital PCR device, the hot bath is an oil bath.
Optionally, in the fully automatic integrated digital PCR device, the thermal cycling assembly further includes a critical low temperature bath and a critical high temperature bath;
the liquid drop slide is circularly switched between the high-temperature bath kettle and the low-temperature bath kettle and further comprises:
after the liquid drop slide is taken out from the low-temperature bath kettle, the liquid drop slide is firstly put into the critical high-temperature bath kettle until the temperature of the liquid drop slide is raised to be close to the first temperature corresponding to the high-temperature bath kettle, and then the liquid drop slide is put into the high-temperature bath kettle for heating;
and after the liquid drop slide is taken out from the high-temperature bath kettle, the liquid drop slide is firstly put into the critical low-temperature bath kettle until the temperature of the liquid drop slide is reduced to be close to the second temperature corresponding to the low-temperature bath kettle, and then the liquid drop slide is put into the high-temperature bath kettle for cooling.
Optionally, in the fully automatic integrated digital PCR apparatus, the temperature range of the critical high-temperature bath is 100 to 150 degrees celsius, inclusive;
the temperature range of the critical low-temperature bath kettle is from-2 ℃ to 20 ℃, including the end points.
Optionally, in the fully automatic integrated digital PCR device, the oil bath medium of the oil bath pan is dimethicone or diphenylsilicone oil.
Optionally, in the fully automatic integrated digital PCR device, the first temperature ranges from 90 degrees celsius to 98 degrees celsius, inclusive;
the second temperature ranges from 50 degrees Celsius to 70 degrees Celsius, inclusive.
Optionally, in the fully automatic integrated digital PCR device, the droplet slide includes a substrate and a flow guide channel;
the flow guide passage is folded back and forth on the substrate.
Optionally, in the fully automatic integrated digital PCR device, the droplet slide is a microfluidic chip.
Optionally, in the fully-automatic integrated digital PCR device, the droplet carrier is a carrier obtained by repeatedly inserting the transparent high polymer hose on the substrate through a spinner or an embroidery machine.
Optionally, in the fully automatic integrated digital PCR device, the transparent high polymer hose is a teflon catheter.
The invention provides a full-automatic integrated digital PCR device, which comprises a liquid drop generating assembly, a thermal circulation drainage channel, a thermal circulation assembly, a fluorescence detection assembly and a controller; the thermal cycle component comprises a liquid drop slide, a mechanical arm, a high-temperature bath kettle and a low-temperature bath kettle; the liquid drops to be detected generated by the liquid drop generating assembly flow into the liquid drop chip through the thermal circulation drainage channel; the mechanical arm is used for circularly switching the liquid drop slide between the high-temperature bath kettle and the low-temperature bath kettle; the fluorescence detection assembly is used for carrying out fluorescence detection on the droplet slide glass after thermal cycling for preset times; the controller is used for controlling the liquid drop generation assembly, the thermal circulation assembly and the fluorescence detection assembly and carrying out quantitative analysis on detection results. According to the invention, the liquid to be detected which needs to be subjected to thermal circulation is placed in the liquid drop slide, then the liquid drop slide is repeatedly transferred in the thermal bath pools with different temperatures through the mechanical arm to realize thermal circulation amplification of nucleic acid, the liquid drop slide is heated in a cladding manner through the thermal bath, so that the liquid drop to be detected is heated more uniformly, meanwhile, the temperature controllability of the thermal bath kettle is better, the thermal circulation component is more stable and efficient, the amplification of the nucleic acid in the finally obtained liquid drop to be detected is closer to the ideal condition in experimental design, and therefore, the detection accuracy is higher.
Drawings
In order to more clearly illustrate the embodiments or technical solutions of the present invention, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained based on these drawings without creative efforts.
FIG. 1 is a schematic structural diagram of an embodiment of a fully automatic integrated digital PCR device according to the present invention;
FIG. 2 is a top view of the thermal cycling assembly of another embodiment of the fully automatic integrated digital PCR device provided by the present invention;
FIG. 3 is a schematic structural diagram of a droplet slide according to still another embodiment of the fully automatic integrated digital PCR device provided by the present invention.
Detailed Description
In order that those skilled in the art will better understand the disclosure, the invention will be described in further detail with reference to the accompanying drawings and specific embodiments. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The core of the invention is to provide a full-automatic integrated digital PCR device, the structure diagram of one embodiment of which is shown in FIG. 1, and is called as the first embodiment, and the device comprises a droplet generation assembly 100, a thermal circulation drainage channel 200, a thermal circulation assembly 300, a fluorescence detection assembly 400 and a controller 500;
the thermal cycle assembly 300 comprises a droplet carrier 302, a mechanical arm 301, a high temperature bath 3031 and a low temperature bath 3034;
the liquid drops to be detected generated by the liquid drop generating assembly 100 flow through the thermal circulation drainage channel 200 and are placed in the liquid drop slide 302;
the mechanical arm 301 is used for circularly switching the droplet carrier sheet 302 between the high-temperature bath 3031 and the low-temperature bath 3034;
the fluorescence detection assembly 400 is used for performing fluorescence detection on the droplet slide 302 after thermal cycling for a preset number of times;
the controller 500 is used for controlling the droplet generation assembly 100, the thermal cycling assembly 300 and the fluorescence detection assembly 400, and performing quantitative analysis on the detection result.
In particular, the hot bath is an oil bath, the oil bath has a wider temperature selection range than a water bath (because the condensation point of oil is lower than that of water and the boiling point of oil is higher than that of water), and the oil is not easy to volatilize and has poor conductivity, so that the normal operation cannot be influenced in case of accidental splashing on other elements of the full-automatic integrated digital PCR device.
The fluorescence detection assembly 400 comprises an excitation light source and an image acquisition unit, wherein the image acquisition unit can be an imaging or light intensity detector such as a CMOS (complementary metal oxide semiconductor), CCD (charge coupled device) or PMT (photomultiplier tube) camera; the excitation light source may be an LED or laser source or other hybrid light source.
The droplet generation assembly 100 includes a syringe pump for generating the separated droplets to be detected, and the syringe pump may be a mechanical syringe pump, a pneumatic pump or a peristaltic pump.
The thermocycling drain 200 can be a Y-catheter or a T-catheter or a cross-catheter.
The first temperature is in a range of 90 degrees Celsius to 98 degrees Celsius, inclusive, such as any of 90.0 degrees Celsius, 95.0 degrees Celsius, or 98.0 degrees Celsius; the second temperature is in a range of 50 degrees celsius to 70 degrees celsius, inclusive, such as any of 50.0 degrees celsius, 60.0 degrees celsius, or 63.0 degrees celsius. The first temperature and the second temperature are high temperature and low temperature in thermal cycle required by nucleic acid amplification in PCR process, the parameter ranges are optimal values obtained after theoretical calculation and actual inspection, and can be adjusted according to actual conditions.
It should be noted that the droplet carrier 302 includes a substrate 3021 and a flow guide path; the flow guide path is folded back and forth on the substrate 3021. In order to ensure that the droplets to be detected simultaneously participate in the thermal cycle as many as possible, the droplet carrier 302 should ensure that the flow guide path is as long as possible, so that the flow guide path is configured to be reciprocally folded back on the substrate 3021, thereby ensuring the length of the flow guide path. It should be noted that the flow guide passage may be a passage formed by a groove on the substrate 3021, or may be a tubular structure independent of the substrate 3021, and may be implemented by a conventional laser engraving numerically controlled lathe. Further, the droplet carrier 302 is a microfluidic chip or a droplet container. The microfluidic chip can be a rectangular chip, a trapezoidal chip, a cylindrical chip or the like.
The full-automatic integrated digital PCR device provided by the invention comprises a liquid drop generating assembly 100, a thermal circulation drainage channel 200, a thermal circulation assembly 300, a fluorescence detection assembly 400 and a controller 500; the thermal cycle assembly 300 comprises a droplet carrier 302, a mechanical arm 301, a high temperature bath 3031 and a low temperature bath 3034; the liquid drops to be detected generated by the liquid drop generating assembly 100 flow through the thermal circulation drainage channel 200 and are placed in the liquid drop slide 302; the mechanical arm 301 is used for circularly switching the droplet carrier sheet 302 between the high-temperature bath 3031 and the low-temperature bath 3034; the fluorescence detection assembly 400 is used for performing fluorescence detection on the droplet slide 302 after thermal cycling for a preset number of times; the controller 500 is used for controlling the droplet generation assembly 100, the thermal cycling assembly 300 and the fluorescence detection assembly 400, and performing quantitative analysis on the detection result. According to the invention, the liquid drop to be detected, which needs to be subjected to thermal circulation, is placed in the liquid drop slide 302, the liquid drop slide 302 is repeatedly transferred in the thermal bath pools with different temperatures through the mechanical arm 301 to realize thermal circulation amplification of nucleic acid, the liquid drop slide 302 is subjected to cladding heating through the thermal bath, so that the liquid drop to be detected is heated more uniformly, meanwhile, the temperature controllability of the thermal bath is better, the thermal circulation component 300 is more stable and efficient, and the amplification of the nucleic acid in the finally obtained liquid drop to be detected is closer to the ideal condition in experimental design, so that the detection accuracy is higher.
On the basis of the first specific embodiment, the hot-bath kettle is further modified to obtain a second specific embodiment, and a schematic structural diagram of the thermal circulation component 300 is shown in fig. 2, and includes a droplet generation component 100, a thermal circulation flow guide 200, a thermal circulation component 300, a fluorescence detection component 400 and a controller 500;
the thermal cycle assembly 300 comprises a droplet carrier 302, a mechanical arm 301, a high temperature bath 3031 and a low temperature bath 3034;
the liquid drops to be detected generated by the liquid drop generating assembly 100 flow through the thermal circulation drainage channel 200 and are placed in the liquid drop slide 302;
the mechanical arm 301 is used for circularly switching the droplet carrier sheet 302 between the high-temperature bath 3031 and the low-temperature bath 3034;
the fluorescence detection assembly 400 is used for performing fluorescence detection on the droplet slide 302 after thermal cycling for a preset number of times;
the controller 500 is configured to control the droplet generation assembly 100, the thermal cycling assembly 300, and the fluorescence detection assembly 400, and perform quantitative analysis on the detection result;
the thermal cycle assembly 300 further comprises a critical low temperature bath 3033 and a critical high temperature bath 3032;
the cyclic switching of the droplet carrier 302 between the high-temperature bath 3031 and the low-temperature bath 3034 further comprises:
after the liquid drop slide glass 302 is taken out from the low-temperature bath 3034, the liquid drop slide glass 302 is firstly put into the critical high-temperature bath 3032 until the temperature of the liquid drop slide glass 302 is raised to be close to the first temperature corresponding to the high-temperature bath 3031, and then the liquid drop slide glass 302 is put into the high-temperature bath 3031 for heating;
after the liquid drop slide glass 302 is taken out from the high-temperature bath 3031, the critical low-temperature bath 3033 is firstly put until the temperature of the liquid drop slide glass 302 is reduced to be close to the second temperature corresponding to the low-temperature bath 3034, and then the liquid drop slide glass 302 is put into the high-temperature bath 3031 for cooling.
The difference between this embodiment and the above embodiment is that the critical low-temperature bath 3033 and the critical high-temperature bath 3032 are added to the hot bath in this embodiment, and the rest of the structure is the same as that in the above embodiment, and therefore, the detailed description thereof is omitted.
In this embodiment, two new thermal baths, namely the critical low-temperature bath 3033 and the critical high-temperature bath 3032, are added, and the droplet carrier 302 needs to perform thermal cycling, namely high-low-temperature-high-temperature-low-temperature cycling, for example, in a process from high temperature to low temperature, since the temperature difference between the high-temperature bath 3031 and the low-temperature bath 3034 is not very large, the droplet carrier 302 needs to be soaked in the low-temperature bath 3034 for a long time to reach the second temperature after being taken out from the high-temperature bath 3031, so that the thermal cycling time is long and the detection efficiency is not high, in this embodiment, the critical low-temperature bath 3033 is introduced, and the temperature difference between the critical low-temperature bath 3033 and the high-temperature bath 3031 is large, so that the droplet carrier 302 can be rapidly cooled, when the droplet carrier 302 is rapidly cooled to the second temperature, and then the reaction product is transferred into the low-temperature bath 3034 for full reaction, thereby greatly accelerating the thermal cycling speed and improving the PCR detection efficiency.
The temperature range of the critical high-temperature bath 3032 is 100 to 150 ℃, including the end points, such as any one of 100.0 ℃, 123.6 ℃ or 150.0 ℃; the temperature range of the critical low-temperature bath 3033 is-20 ℃ to 20 ℃, inclusive, such as any one of-20.0 ℃, 12.0 ℃ or 20.0 ℃, wherein-20 ℃ means twenty ℃ below zero.
Fig. 2 is a top view of the thermal cycle assembly 300.
On the basis of the second embodiment, the droplet carrier 302 is further improved to obtain the third embodiment, and the structural schematic diagram of the droplet carrier 302 is shown in fig. 3, and includes a droplet generation assembly 100, a thermal circulation flow guide 200, a thermal circulation assembly 300, a fluorescence detection assembly 400, and a controller 500;
the thermal cycle assembly 300 comprises a droplet carrier 302, a mechanical arm 301, a high temperature bath 3031 and a low temperature bath 3034;
the liquid drops to be detected generated by the liquid drop generating assembly 100 flow through the thermal circulation drainage channel 200 and are placed in the liquid drop slide 302;
the mechanical arm 301 is used for circularly switching the droplet carrier sheet 302 between the high-temperature bath 3031 and the low-temperature bath 3034;
the fluorescence detection assembly 400 is used for performing fluorescence detection on the droplet slide 302 after thermal cycling for a preset number of times;
the controller 500 is configured to control the droplet generation assembly 100, the thermal cycling assembly 300, and the fluorescence detection assembly 400, and perform quantitative analysis on the detection result;
the thermal cycle assembly 300 further comprises a critical low temperature bath 3033 and a critical high temperature bath 3032;
the cyclic switching of the droplet carrier 302 between the high-temperature bath 3031 and the low-temperature bath 3034 further comprises:
after the liquid drop slide glass 302 is taken out from the low-temperature bath 3034, the liquid drop slide glass 302 is firstly put into the critical high-temperature bath 3032 until the temperature of the liquid drop slide glass 302 is raised to be close to the first temperature corresponding to the high-temperature bath 3031, and then the liquid drop slide glass 302 is put into the high-temperature bath 3031 for heating;
after the liquid drop slide glass 302 is taken out from the high-temperature bath 3031, the liquid drop slide glass 302 is firstly put into the critical low-temperature bath 3033 until the temperature of the liquid drop slide glass 302 is reduced to be close to the second temperature corresponding to the low-temperature bath 3034, and then the liquid drop slide glass 302 is put into the high-temperature bath 3031 for cooling;
the droplet carrier 302 is a carrier obtained by repeatedly inserting the transparent polymer hose 3022 on the substrate 3021 through a spinner or an embroidery machine.
The difference between this embodiment and the above embodiments is that the shape and obtaining manner of the droplet carrier 302 are defined in this embodiment, and the rest of the structure is the same as that of the above embodiments, and will not be described herein again.
Furthermore, the transparent polymer hose 3022 is a teflon conduit, which has excellent toughness and softness and can be used as a wire on a cloth spinner, and the teflon conduit has high transparency, so that subsequent fluorescence monitoring is not affected, and the measurement accuracy can be further improved.
In the specific embodiment, the transparent high polymer conduit is sewn on the substrate 3021 by using a spinner or an embroidery machine to prepare the droplet carrier 302, so that the production equipment is low in cost, the raw materials of the equipment are easily available, the production requirement of the droplet carrier 302 is greatly reduced, the universality of the invention is expanded, and the production cost of the full-automatic integrated digital PCR device is greatly reduced.
It should be noted that, in fig. 3, the left side is a schematic view of a spinning machine, and the right side is a schematic view of the droplet carrier 302; the black horizontal line on the transparent polymer tubing 3022 indicates where the transparent polymer tubing 3022 passes through the substrate to the other side.
The embodiments are described in a progressive manner, each embodiment focuses on differences from other embodiments, and the same or similar parts among the embodiments are referred to each other. The device disclosed by the embodiment corresponds to the method disclosed by the embodiment, so that the description is simple, and the relevant points can be referred to the method part for description.
It is to be noted that, in the present specification, relational terms such as first and second, and the like are used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other identical elements in a process, method, article, or apparatus that comprises the element.
The fully automatic integrated digital PCR device provided by the invention is described in detail above. The principles and embodiments of the present invention are explained herein using specific examples, which are presented only to assist in understanding the method and its core concepts. It should be noted that, for those skilled in the art, it is possible to make various improvements and modifications to the present invention without departing from the principle of the present invention, and those improvements and modifications also fall within the scope of the claims of the present invention.
Claims (10)
1. A full-automatic integrated digital PCR device is characterized by comprising a liquid drop generating assembly, a thermal circulation drainage channel, a thermal circulation assembly, a fluorescence detection assembly and a controller;
the thermal cycle component comprises a liquid drop slide, a mechanical arm, a high-temperature bath kettle and a low-temperature bath kettle;
the liquid drops to be detected generated by the liquid drop generating assembly flow into the liquid drop chip through the thermal circulation drainage channel;
the mechanical arm is used for circularly switching the liquid drop slide between the high-temperature bath kettle and the low-temperature bath kettle;
the fluorescence detection assembly is used for carrying out fluorescence detection on the droplet slide glass after thermal cycling for preset times;
the controller is used for controlling the liquid drop generation assembly, the thermal circulation assembly and the fluorescence detection assembly and carrying out quantitative analysis on detection results.
2. The fully automated integrated digital PCR device according to claim 1, wherein the thermal cycling assembly further comprises a critical low temperature bath and a critical high temperature bath;
the liquid drop slide is circularly switched between the high-temperature bath kettle and the low-temperature bath kettle and further comprises:
after the liquid drop slide is taken out from the low-temperature bath kettle, the liquid drop slide is firstly put into the critical high-temperature bath kettle until the temperature of the liquid drop slide is raised to be close to the first temperature corresponding to the high-temperature bath kettle, and then the liquid drop slide is put into the high-temperature bath kettle for heating;
and after the liquid drop slide is taken out from the high-temperature bath kettle, the liquid drop slide is firstly put into the critical low-temperature bath kettle until the temperature of the liquid drop slide is reduced to be close to the second temperature corresponding to the low-temperature bath kettle, and then the liquid drop slide is put into the high-temperature bath kettle for cooling.
3. The fully automated integrated digital PCR device according to claim 1, wherein the hot bath is an oil bath.
4. The fully automatic integrated digital PCR device according to claim 3, wherein the critical high temperature bath has a temperature range of 100 to 150 degrees celsius, inclusive;
the critical low temperature bath temperature ranges from-20 ℃ to 20 ℃, inclusive.
5. The fully automatic integrated digital PCR device according to claim 4, wherein the oil bath medium of the oil bath pan is dimethicone or diphenylsilicone oil.
6. The fully automated, integrated digital PCR device according to claim 3, wherein the first temperature ranges from 90 degrees celsius to 98 degrees celsius, inclusive;
the second temperature ranges from 50 degrees Celsius to 70 degrees Celsius, inclusive.
7. The fully automatic integrated digital PCR device according to claim 1, wherein the droplet slide comprises a substrate and a flow guide path;
the flow guide passage is folded back and forth on the substrate.
8. The fully automatic integrated digital PCR device according to claim 7, wherein the droplet slides are microfluidic chips.
9. The fully automatic integrated digital PCR device according to claim 7, wherein the droplet carrier is a carrier obtained by repeatedly inserting a transparent high polymer hose on the substrate through a spinner or an embroidery machine.
10. The fully automated integrated digital PCR device according to claim 9, wherein the transparent polymer hose is a teflon catheter.
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| CN111790462A (en) * | 2020-08-13 | 2020-10-20 | 河南固创生物科技有限公司 | Heating device for micro-fluidic detection |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2181509Y (en) * | 1993-12-02 | 1994-11-02 | 李侃谆 | Mechanical arm DNA augmentor with curved track running mode |
| JP2008278791A (en) * | 2007-05-10 | 2008-11-20 | Tokyo Medical & Dental Univ | Nucleic acid amplification apparatus, method, cell culturing, and nucleic acid amplification method |
| US8741570B2 (en) * | 2008-02-06 | 2014-06-03 | Ludwig-Maximilians-Universitat Munchen | Thermo-optical characterisation of nucleic acid molecules |
| CN105505761A (en) * | 2015-12-21 | 2016-04-20 | 中国科学院苏州生物医学工程技术研究所 | Digital isothermal nucleic acid detecting device and detecting method thereof |
| CN107164523A (en) * | 2017-06-27 | 2017-09-15 | 中国科学院长春光学精密机械与物理研究所 | A kind of method and device for digital pcr |
| CN108654709A (en) * | 2018-05-08 | 2018-10-16 | 中国科学院苏州生物医学工程技术研究所 | A kind of micro-fluidic chip and its method of manufacturing technology |
| CN109266516A (en) * | 2018-09-28 | 2019-01-25 | 中国科学院长春光学精密机械与物理研究所 | The production method and detection device of DNA cloning device, DNA cloning device |
| CN109562383A (en) * | 2016-06-10 | 2019-04-02 | 星阵私人有限公司 | Rapid thermal cycles for sample analysis and processing |
| CN110066859A (en) * | 2018-01-24 | 2019-07-30 | 思纳福(北京)医疗科技有限公司 | Digital pcr detector |
| CN110591911A (en) * | 2019-09-30 | 2019-12-20 | 中国科学院长春光学精密机械与物理研究所 | Optical-mechanical-electrical integrated full-automatic digital PCR device |
-
2019
- 2019-12-06 CN CN201911241203.9A patent/CN110846206A/en active Pending
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2181509Y (en) * | 1993-12-02 | 1994-11-02 | 李侃谆 | Mechanical arm DNA augmentor with curved track running mode |
| JP2008278791A (en) * | 2007-05-10 | 2008-11-20 | Tokyo Medical & Dental Univ | Nucleic acid amplification apparatus, method, cell culturing, and nucleic acid amplification method |
| US8741570B2 (en) * | 2008-02-06 | 2014-06-03 | Ludwig-Maximilians-Universitat Munchen | Thermo-optical characterisation of nucleic acid molecules |
| CN105505761A (en) * | 2015-12-21 | 2016-04-20 | 中国科学院苏州生物医学工程技术研究所 | Digital isothermal nucleic acid detecting device and detecting method thereof |
| CN109562383A (en) * | 2016-06-10 | 2019-04-02 | 星阵私人有限公司 | Rapid thermal cycles for sample analysis and processing |
| CN107164523A (en) * | 2017-06-27 | 2017-09-15 | 中国科学院长春光学精密机械与物理研究所 | A kind of method and device for digital pcr |
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| CN109266516A (en) * | 2018-09-28 | 2019-01-25 | 中国科学院长春光学精密机械与物理研究所 | The production method and detection device of DNA cloning device, DNA cloning device |
| CN110591911A (en) * | 2019-09-30 | 2019-12-20 | 中国科学院长春光学精密机械与物理研究所 | Optical-mechanical-electrical integrated full-automatic digital PCR device |
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