CN104946511A - Laminated structure chamber internal temperature sensing PCR chip - Google Patents
Laminated structure chamber internal temperature sensing PCR chip Download PDFInfo
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- CN104946511A CN104946511A CN201510290092.6A CN201510290092A CN104946511A CN 104946511 A CN104946511 A CN 104946511A CN 201510290092 A CN201510290092 A CN 201510290092A CN 104946511 A CN104946511 A CN 104946511A
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- electrode
- reaction chamber
- pcr chip
- temperature sensing
- substrate
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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
- 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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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
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- Molecular Biology (AREA)
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- Automatic Analysis And Handling Materials Therefor (AREA)
Abstract
The invention provides a laminated structure chamber internal temperature sensing PCR chip. The laminated structure chamber internal temperature sensing PCR chip comprises an upper substrate, a temperature measuring electrode, a reaction chamber, a lower substrate and a heating electrode which are sequentially arranged from top to bottom, wherein the temperature measuring electrode is deposited on one side of the substrate; the reaction chamber is etched on the upper surface of the lower substrate, and the reaction chamber is used for containing a reaction solution; the heating electrode is deposited on the lower surface of the lower substrate; the temperature measuring electrode deposited side of the upper substrate is combined with the reaction chamber etched side of the lower substrate to form an integral body; and the left side and the right side of the reaction chamber are respectively provided with a sample inlet and a sample outlet in order to realize direct connection with the outside. The PCR chip has the advantages of small size, small amount of consumed reagents, reduction of the liquid overflow and bubble generation in the high temperature reaction process due to adoption of glass as the substrate, realization of small non-heating area and uniform temperature distribution due to uniform distribution of the electrode, and realization of direct and accurate measurement of the liquid temperature due to stretching of the temperature measuring electrode to the reaction solution.
Description
Technical field
The present invention relates to the pcr chip of PCR biological technical field, particularly, relate to temperature sensing pcr chip in a kind of rhythmo structure chamber.
Background technology
Polymerase chain reaction PCR is that a kind of external high speed conventional in bio-science and medical field increases the Protocols in Molecular Biology of specific DNA fragments in a large number.In PCR, a circulation is divided into three sections of regions, high-temperature denatured (about 95 DEG C), low-temperature annealing (about 55 DEG C), middle temperature extend (about 70 DEG C), on the PCR instrument device of routine, a circulation approximately needs about 3min, and during this period of time in have quite a few for the lifting of temperature, time availability is lower.The principal element affecting PCR reaction success or failure and amplification efficiency comprises temperature homogeneity, temperature-controlled precision, temperature rate.Existing Standard PCR chip, amplification efficiency is low, and the micro-fluidic pcr chip adopting MEMS technology to realize exists the problems such as temperature homogeneity is bad, thermometric is inaccurate.
Summary of the invention
For defect of the prior art, the object of this invention is to provide temperature sensing pcr chip in a kind of rhythmo structure chamber, directly can measure reacting liquid temperature, reduce reactant structure simultaneously, reduce the consumption of the energy and reagent, improve temperature homogeneity and temperature rate.
For realizing above object, the invention provides temperature sensing pcr chip in a kind of rhythmo structure chamber, comprise upper substrate, thermometric electrode, reaction chamber, lower substrate, the heating electrode of arranging successively from top to bottom, wherein: thermometric electrodeposition closes on the one side of reaction chamber in upper substrate; Reaction chamber etches the upper surface in lower substrate, and reaction chamber is used for holding reaction solution; Heating electrode is deposited on the lower surface of lower substrate; The upper substrate one side the depositing thermometric electrode shape that combines with the lower substrate one side being etched with reaction chamber is in aggregates; The left and right sides of reaction chamber is respectively arranged with injection port and is directly connected with the external world with outlet, and injection port notes sample before reacting, and outlet is for reacting rear draw samples.
Preferably, described thermometric electrode extend into reaction chamber inside and contacts with the reaction solution in reaction chamber, directly measures reacting liquid temperature.
Preferably, the material of described upper substrate and lower substrate is glass, and the junction of upper substrate and lower substrate is airtight by substrate bonding layer.
More preferably, the material of described substrate bonding layer is PDMS material.
Preferably, described thermometric electrode surface deposition thermometric electrode protecting layer, after thermometric electrode stretches into reaction solution, thermometric electrode protecting layer makes thermometric electrode and reaction solution isolate, and prevents thermometric electrode to be destroyed.
More preferably, the material of described thermometric protective layer is earth silicon material.
Preferably, described thermometric electrode has solder joint to be exposed to outward so that be connected with peripheral circuit.
Preferably, described reaction chamber is the hexagonal structure without upper cover.
Preferably, the area of described heating electrode is greater than reaction chamber, to ensure whole reaction chamber homogeneous temperature.
Preferably, described heating electrode adopts serpentine configuration, and heating electrode section width is greater than 10:1, to ensure temperature homogeneity with gap width ratio.
Preferably, described heating electrode deposits one deck heating electrode protective layer.
Compared with prior art, the present invention has following beneficial effect:
Pcr chip volume of the present invention is little, consumes reagent less, does substrate, can reduce the spilling of liquid and the generation of bubble during pyroreaction with glass; Distribution of electrodes is even, and non-heated region is little, temperature distribution evenly; Thermometric electrode stretches in reaction solution, can directly measure the temperature of a liquid, and it is more accurate to measure.
Accompanying drawing explanation
By reading the detailed description done non-limiting example with reference to the following drawings, other features, objects and advantages of the present invention will become more obvious:
Fig. 1 is the overall diagram of temperature sensing pcr chip in rhythmo structure chamber of the present invention;
Fig. 2 is the vertical view of temperature sensing pcr chip in rhythmo structure chamber of the present invention;
Fig. 3 is the upward view of temperature sensing pcr chip in rhythmo structure chamber of the present invention;
Fig. 4 is the sectional view of temperature sensing pcr chip in rhythmo structure chamber of the present invention;
Fig. 5 is the temperature control system functional block diagram of temperature sensing pcr chip in rhythmo structure chamber of the present invention;
In figure: substrate on 1,2 substrate bonding layer, 3 thermometric electrodes, 4 thermometric electrode protecting layers, 5 reaction chambers, 6 times substrates, 7 heating electrodes, 8 heating electrode protective layers, 9 injection ports, 10 outlets.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in detail.Following examples will contribute to those skilled in the art and understand the present invention further, but not limit the present invention in any form.It should be pointed out that to those skilled in the art, without departing from the inventive concept of the premise, some distortion and improvement can also be made.These all belong to protection scope of the present invention.
As shown in Figure 1,2,3, 4, the present embodiment provides temperature sensing pcr chip in a kind of rhythmo structure chamber, comprise upper substrate 1, thermometric electrode 3, reaction chamber 5, lower substrate 6, the heating electrode 7 of arranging successively from top to bottom, wherein: thermometric electrode 3 be deposited on substrate 1 close on reaction chamber 5 one side; Reaction chamber 5 etches the upper surface in lower substrate 6, and reaction chamber 5 is for holding reaction solution; Heating electrode 7 is deposited on the lower surface of lower substrate 6; Upper substrate 1 one side depositing thermometric electrode 3 and the lower substrate 6 one side shape that combines being etched with reaction chamber 5 in aggregates; The left and right sides of reaction chamber 5 is respectively arranged with injection port 9 and is directly connected with the external world with outlet 10, and injection port 9 notes sample before reacting, and outlet 10 is for reacting rear draw samples.
As one preferred embodiment, it is inner that described thermometric electrode 3 extend into reaction chamber 5, directly contacts with the reaction solution in reaction chamber 5, directly measure reacting liquid temperature.
As one preferred embodiment, described upper substrate 1 and the junction of lower substrate 6 are by substrate bonding layer 2 airtight (as shown in Figure 4).
As one preferred embodiment, the material of described substrate bonding layer 2 is PDMS material.
As one preferred embodiment, described thermometric electrode 3 surface deposition thermometric electrode protecting layer 4 (as shown in Figure 4), after thermometric electrode 3 stretches into reaction solution, thermometric electrode protecting layer 4 makes thermometric electrode 3 and reaction solution isolate, and prevents thermometric electrode 3 to be destroyed.
As one preferred embodiment, the material of described thermometric protective layer 4 is silicon-dioxide.
As one preferred embodiment, described thermometric electrode 3 has solder joint to be exposed to outward so that be connected with peripheral circuit.
As one preferred embodiment, described reaction chamber 5 is the hexagonal structure (as shown in Figure 1, 2, 3) without upper cover.
As one preferred embodiment, the area of described heating electrode 7 is greater than reaction chamber 5, to ensure whole reaction chamber 5 homogeneous temperature.
As one preferred embodiment, described heating electrode 7 adopts serpentine configuration (as shown in Figure 3), and heating electrode 7 sections of width are greater than 10:1, to ensure temperature homogeneity with gap width ratio.
As one preferred embodiment; described heating electrode 7 deposits one deck heating electrode protective layer 8; thermometric electrode 3 upper surface deposits thermometric electrode protecting layer 4 (as shown in Figure 4); material is silicon-dioxide; not oxidized in work or put procedure for the protection of heating electrode 7 and thermometric electrode 3, and prevent machinery and scratch or the corrosion of other chemical substance.
Be illustrated in figure 5 temperature control system functional block diagram of the present invention, first on human-computer interaction interface, set the parameter needed in reaction, then start sequence of control, chip is started working.By temperature sensing unit, the temperature data collected is transferred to data collecting card, sends actual temperature to human-computer interaction interface and show.Meanwhile, temperature control modules judges the difference of Current Temperatures and target temperature, sends heating or cooling signal to data collecting card, starts heating or cooling execution module, finally makes actual temperature and target temperature be consistent.
Above specific embodiments of the invention are described.It is to be appreciated that the present invention is not limited to above-mentioned particular implementation, those skilled in the art can make various distortion or amendment within the scope of the claims, and this does not affect flesh and blood of the present invention.
Claims (10)
1. a temperature sensing pcr chip in rhythmo structure chamber, is characterized in that, comprises upper substrate, thermometric electrode, reaction chamber, lower substrate, the heating electrode of arranging successively from top to bottom, wherein: thermometric electrodeposition is in the one side of substrate; Reaction chamber etches the upper surface in lower substrate, and reaction chamber is used for holding reaction solution; Heating electrode is deposited on the lower surface of lower substrate; The upper substrate one side the depositing thermometric electrode shape that combines with the lower substrate one side being etched with reaction chamber is in aggregates; The left and right sides of reaction chamber is respectively arranged with injection port and is directly connected with the external world with outlet, and injection port notes sample before reacting, and outlet is for reacting rear draw samples.
2. temperature sensing pcr chip in a kind of rhythmo structure chamber according to claim 1, is characterized in that, the material of described upper substrate and lower substrate is glass, and the junction of upper substrate and lower substrate is airtight by substrate bonding layer.
3. temperature sensing pcr chip in a kind of rhythmo structure chamber according to claim 2, is characterized in that, the material of described substrate bonding layer is PDMS material.
4. temperature sensing pcr chip in a kind of rhythmo structure chamber according to claim 1, is characterized in that, described thermometric electrode extend into reaction chamber inside and contacts with the reaction solution in reaction chamber, directly measures reacting liquid temperature.
5. temperature sensing pcr chip in a kind of rhythmo structure chamber according to claim 4; it is characterized in that; described thermometric electrode surface deposition thermometric electrode protecting layer, after thermometric electrode stretches into reaction solution, thermometric electrode protecting layer makes thermometric electrode and reaction solution isolate, and prevents thermometric electrode to be destroyed.
6. temperature sensing pcr chip in a kind of rhythmo structure chamber according to claim 5, is characterized in that, the material of described thermometric protective layer is silicon-dioxide.
7. temperature sensing pcr chip in a kind of rhythmo structure chamber according to any one of claim 1-6, is characterized in that, it is outer so that be connected with peripheral circuit that described thermometric electrode has solder joint to be exposed to.
8. temperature sensing pcr chip in a kind of rhythmo structure chamber according to any one of claim 1-6, is characterized in that, described reaction chamber is the hexagonal structure without upper cover.
9. temperature sensing pcr chip in a kind of rhythmo structure chamber according to any one of claim 1-6, it is characterized in that, the area of described heating electrode is greater than reaction chamber, to ensure whole reaction chamber homogeneous temperature.
10. temperature sensing pcr chip in a kind of rhythmo structure chamber according to any one of claim 1-6, is characterized in that, described heating electrode adopts serpentine configuration, and heating electrode section width is greater than 10:1, to ensure temperature homogeneity with gap width ratio; Described heating electrode deposits one deck heating electrode protective layer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201510290092.6A CN104946511A (en) | 2015-05-29 | 2015-05-29 | Laminated structure chamber internal temperature sensing PCR chip |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510290092.6A CN104946511A (en) | 2015-05-29 | 2015-05-29 | Laminated structure chamber internal temperature sensing PCR chip |
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| CN104946511A true CN104946511A (en) | 2015-09-30 |
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| CN201510290092.6A Pending CN104946511A (en) | 2015-05-29 | 2015-05-29 | Laminated structure chamber internal temperature sensing PCR chip |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107723227A (en) * | 2016-08-12 | 2018-02-23 | 广州康昕瑞基因健康科技有限公司 | Gene sequencing reaction cell and gene sequencing reaction device |
| CN109632721A (en) * | 2018-12-28 | 2019-04-16 | 中国科学院电子学研究所 | A kind of LRSPR- fluorescence imaging parallel detection device and LRSPR chip manufacture method |
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| CN102929309A (en) * | 2005-01-25 | 2013-02-13 | 欧西里其有限责任公司 | Temperature controller for small fluid samples having different heat capacities |
| CN103103120A (en) * | 2013-01-18 | 2013-05-15 | 上海交通大学 | Integrated temperature controlled PCR-CE (polymerase chain reaction-capillary electrophoresis) micro-fluidic chip and preparation method thereof |
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2015
- 2015-05-29 CN CN201510290092.6A patent/CN104946511A/en active Pending
Patent Citations (6)
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| CN1368555A (en) * | 2001-02-09 | 2002-09-11 | 中国科学院电子学研究所 | DNA-PCR biochip and miniature heat circulator using it |
| CN102929309A (en) * | 2005-01-25 | 2013-02-13 | 欧西里其有限责任公司 | Temperature controller for small fluid samples having different heat capacities |
| CN102083533A (en) * | 2008-04-10 | 2011-06-01 | 芬兰技术研究中心 | Microfluidic chip devices and their use |
| KR20100124453A (en) * | 2009-05-19 | 2010-11-29 | 경북대학교 산학협력단 | Pcr chamber chip and method for manufacturing pcr chamber chip |
| CN103103120A (en) * | 2013-01-18 | 2013-05-15 | 上海交通大学 | Integrated temperature controlled PCR-CE (polymerase chain reaction-capillary electrophoresis) micro-fluidic chip and preparation method thereof |
| CN104593256A (en) * | 2015-01-06 | 2015-05-06 | 上海交通大学 | PCR chip with repeatedly used electrode |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107723227A (en) * | 2016-08-12 | 2018-02-23 | 广州康昕瑞基因健康科技有限公司 | Gene sequencing reaction cell and gene sequencing reaction device |
| CN109632721A (en) * | 2018-12-28 | 2019-04-16 | 中国科学院电子学研究所 | A kind of LRSPR- fluorescence imaging parallel detection device and LRSPR chip manufacture method |
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Application publication date: 20150930 |