CN105665046A - Fast-temperature-rise microfluidic chip system - Google Patents
Fast-temperature-rise microfluidic chip system Download PDFInfo
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- CN105665046A CN105665046A CN201610116013.4A CN201610116013A CN105665046A CN 105665046 A CN105665046 A CN 105665046A CN 201610116013 A CN201610116013 A CN 201610116013A CN 105665046 A CN105665046 A CN 105665046A
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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
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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
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N23/00—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00
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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
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
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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
- B01L2300/00—Additional constructional details
- B01L2300/18—Means for temperature control
- B01L2300/1805—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
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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
- B01L2300/00—Additional constructional details
- B01L2300/18—Means for temperature control
- B01L2300/1894—Cooling means; Cryo cooling
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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
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
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Abstract
The invention relates to a fast-temperature-rise microfluidic chip system, which comprises a chip, a support frame, a two-dimensional regulating platform and an injection pump, wherein the chip comprises a substrate, a cover sheet, an electromagnetic valve, a refrigeration device and a heating device; the substrate is provided with a microfluidic passage; a constant-temperature liquid storage region, a temperature jumping region and a test region are sequentially formed in the microfluidic passage; the cover sheet covers and seals the substrate; the electromagnetic valve is communicated with the microfluidic passage; the refrigeration device is arranged in a way of being tightly attached to the constant-temperature liquid storage region of the substrate; the heating device is arranged in a way of being tightly attached to the temperature jumping region of the substrate; the test region is exposed in the irradiation of synchronous radiation light for performing test; the support frame is used for installing and fixing the chip; the two-dimensional regulating platform is used for regulating the position of the support frame and comprises a horizontal position regulating module and a vertical position regulating module; the injection pump is provided with a body and a first valve opening connected with the body; the valve opening is communicated with the microfluidic passage of the chip. The chip system provided by the invention can realize the fast temperature pumping of a solution sample, and is used for synchronous radiation reaction dynamic study.
Description
Technical field
The present invention relates to a kind of chip system for studying kinetics, relate more specifically to a kind of fast temperature combined with synchrotron radiation measurement and rise to micro flow chip system.
Background technology
The research of kinetics is the main path of research reaction mechanism. Reaction half-time is shorter than to the fast response of 1s, adopts conventional analytical techniques cannot reaction be studied. Kinetics has multiple research method, including continuous stream/stop-flow technique, temperature-jump Relaxation Method, flash photolysis technology etc. Wherein, temperature-jump Relaxation Method is a kind of common experimental technology studying fast response in solution, namely within the of short duration time, equilibrium system is applied a thermal perturbation, and in conjunction with detection equipment, the systems such as such as chromatograph, mass spectrum, nuclear magnetic resonance, NMR and absorption spectrum, monitor reaction process by measuring a certain characteristic of solution. Therefore, the design of temperature-jump relaxation device it is crucial that choose one to be brought rapidly up scheme with detection means matches.
Synchrotron Radiation is to utilize electronics to do high-speed motion in magnetic field to produce the large-scale scientific experiments device of synchrotron radiation light. Synchrotron radiation light source is a kind of new type light source, covers X ray, VUV, visible ray to far infrared band, and continuously adjustabe. Synchrotron radiation light source has high well-illuminated, high-space resolution, high time resolution feature, is detecting material structure and the parsing strong instrument of reaction mechanism.
At present, the method producing temperature-jump mainly has capacitor discharge, microwave and pulse laser. Utilizing capacitor discharge method to produce Temperature jump, it is necessary to higher salinity and ionic strength, and conducting solution need to add the voltage of thousands of volts, this will cause the adverse effects such as the polarization of molecule; Microwave method is only applicable to polar solvent; Pulsed laser deposition need to consider the absorbing medium light sensitivity in a certain optical maser wavelength. Considering the complexity of said temperature inshot, if combining with synchrotron radiation light source detection means, in addition it is also necessary to require and layout according to concrete line station, carrying out software and the transformation of hardware many-side, comparatively inconvenience.
In recent years, synchrotron radiation is increasingly becoming kinetics study hotspot in conjunction with micro-fluidic mixed type array experiment technology.Basic operation unit involved in biological and chemical field is integrated on the chip of several square centimeters by micro-fluidic chip, microchannel form network, run through whole system with controlled fluid. Utilize the Laminar flow mixing mechanism under the dynamic focus hybrid mode of microfluid or low reynolds number, reactant is quickly propelled in micro flow chip and mixes rapidly, with synchrotron radiation light for probe, it is achieved kinetics dynamic studies. Synchrotron radiation is convenient in conjunction with micro flow chip experimental system composition, cost is low, functional integration is high.
But, up to the present, micro flow chip is used for the research of synchrotron radiation kinetics mainly by the flow behavior of fluid under minute yardstick so that solution quickly mixes, and is mainly used in continuous stream/stop-flow technique. And utilize fluidic heat exchange of fluids in microchannel to be significantly stronger than conventional yardstick and change thermal property, there is not been reported in the research that is applied to temperature-jump relaxation of being combined with synchrotron radiation by micro flow chip.
Summary of the invention
It is desirable to provide a kind of fast temperature rises to micro flow chip system, this system measures technology in combinations with synchrotron radiation X-ray, for the kinetics correlational study of temperature-jump relaxation.
Fast temperature of the present invention rises to micro flow chip system, including: chip, this chip includes substrate, cover plate, electromagnetic valve, refrigerating plant and heater, substrate has microchannel, this microchannel sequentially forms constant temperature liquid storage district, Temperature jump district and test section, cover plate covers and seal substrate, electromagnetic valve connection microchannel, refrigerating plant is close to the constant temperature liquid storage district of substrate and is arranged, heater is close to the Temperature jump district of substrate and is arranged, and test section is exposed under the irradiation of synchrotron radiation light and tests; For installing the support fixing this chip; For regulating the two-dimension adjustment platform of the position of support, this two-dimension adjustment platform includes horizontal level adjustment module and upright position adjustment module; And syringe pump, this syringe pump has body and the first valve port being connected with this body, and described valve port connects with the microchannel of chip.
Microchannel includes relatively independent the first microchannel, the second microchannel and the 3rd microchannel, wherein, the arrival end of the first microchannel is connected with syringe pump, the port of export of the first microchannel is connected with electromagnetic valve, the arrival end of the second microchannel is connected with sample liquids, the port of export of the second microchannel is connected with electromagnetic valve, and the arrival end of the 3rd microchannel is connected with electromagnetic valve.
Microchannel also includes the 4th microchannel and the 5th microchannel that are set in turn in the 3rd downstream, microchannel, the 4th formation temperature saltus step district, microchannel, the 5th formation test section, microchannel.
4th microchannel is that width first disperses the microchannel restrained afterwards. Preferably, the 4th microchannel is rectangle-trapezoidal microchannel.
With first, second, third and the 5th compared with microchannel, the width of the 4th microchannel is relatively big and the degree of depth is less.
This chip also includes the warming probe at the test position place being arranged at the first microchannel.
This syringe pump also includes the second valve port with air communication.
This syringe pump also includes the 3rd valve port connected with cleanout fluid.
This support includes base plate and is perpendicular to the installing plate of this base plate, and this installing plate has through hole, and chip is fixed in this through hole.
This heater is the Pt electrode of sputtering patterning on the glass substrate.
Micro flow chip system is risen to by the fast temperature of the present invention, sample solution can carry out constant temperature pretreatment in constant temperature liquid storage district, after temperature constant, sample can be quickly propelled to Temperature jump district, sample temperature rises rapidly, test subsequently into synchrotron radiation X-ray test section, owing to sample takes to flow to continuously sample loading mode, namely after sample occurrence temperature rises to, the distance that the length in response time flows through in microchannel to sample is directly proportional, in different distances, the multidate information of sample is detected by synchrotron radiation X-ray, react dynamic (dynamical) research.Fast temperature according to the present invention rises to micro flow chip system and also has the advantages that composition is convenient, simple to operate; Utilize the advantage that micro-fluidic chip function is integrated, under the accurately control of computer, syringe pump, electromagnetic valve, refrigerating plant, heater etc. cooperate, it is achieved that solution example auto injection, constant temperature pretreatment, the function such as microchannel cleans automatically, dry; Chip can improve upgrading according to experimental result and freely replace to other chips, and experimental system miscellaneous part is all reusable, reduces processing and manufacturing cost; The control of this system and data acquisition interface are based on synchrotron radiation light source control system platform and write, the correlation control unit and the driver that adopt are that synchrotron radiation is special, meet real needs and the layout of synchrotron radiation station, become one with experiment centre miscellaneous equipment.
Accompanying drawing explanation
Fig. 1 is the schematic diagram that fast temperature according to a preferred embodiment of the present invention rises to micro flow chip system;
Fig. 2 is the overall structure schematic diagram of the support of Fig. 1;
Fig. 3 illustrates the overall structure of the rack-mount chip of Fig. 1;
Fig. 4 is the perspective view of the cover plate of Fig. 3;
Fig. 5 is the top view of the substrate of Fig. 3.
Detailed description of the invention
Below in conjunction with accompanying drawing, provide presently preferred embodiments of the present invention, and be described in detail.
Fig. 1 is the schematic diagram that fast temperature according to a preferred embodiment of the present invention rises to micro flow chip system, and this fast temperature rises to micro flow chip system 1 and includes computer 11, two-dimension adjustment platform 12, syringe pump 13, fluid supply 14, support 15 and chip 16. Wherein, chip 16 is fixed in support 15, and support 15 is arranged on two-dimension adjustment platform 12, adjusts the particular location of chip 16 thereby through two-dimension adjustment platform 12. Syringe pump 13 is connected with chip 16, and computer 11 is connected with two-dimension adjustment platform 12, syringe pump 13 and chip 16 communication respectively.
Two-dimension adjustment platform 12 includes horizontal level adjustment module 121 and upright position adjustment module 122. Support 15 is fixed in upright position adjustment module 122, and upright position adjustment module 122 is fixed in horizontal level adjustment module 121, this horizontal level adjustment module 121 and upright position adjustment module 122 are connected with computer 11 communication, horizontal level adjustment module 121 and upright position adjustment module 122 is controlled thereby through computer 11, thus accurately controlling the particular location of support 15, thus accurately regulate the chip 16 position relative to synchrotron radiation hot spot.
Syringe pump 13 includes body 131 and the first valve port the 132, second valve port the 133, the 3rd valve port 134 and the 4th valve port 135 being connected with body. Wherein, the first valve port 132 is connected by conduit with chip 16, the second valve port 133 and air communication, and the 3rd valve port 134 connects with fluid supply 14 respectively with the 4th valve port 135. In operation, this syringe pump 13 receives the signal behavior valve port of computer 11, arranges sampling volume and speed, it is achieved Liquid sample introduction, microchannel clean and the function such as dry.
Fluid supply 14 includes holding the first container 141 of sample solution, holds the second container 142 of waste liquid, hold the 3rd container 143 of the first cleanout fluid and hold the 4th container 144 of the second cleanout fluid, wherein, first container 141 is connected with chip 16 respectively with second container 142, and the 3rd container 143 is connected with the 3rd valve port 134 of syringe pump 13 and the 4th valve port 135 respectively with the 4th container 144.
Fig. 2 is the overall structure schematic diagram that fast temperature according to a preferred embodiment of the present invention rises to the support of micro flow chip system, this support 15 is T-shaped, it includes base plate 151 and is perpendicular to the installing plate 152 of this base plate 151, the center of this installing plate 152 has through hole 152a, chip 16 is fixed in this through hole 152a, as shown in Figure 3. The base of this base plate 151 is provided with screw hole, consequently facilitating be fixed in upright position adjustment module 122 by support 15.
Fig. 3 illustrates that the overall structure of rack-mount chip, chip 16 include substrate 161, cover plate 162, electromagnetic valve 163, refrigerating plant 164 and heater 165. Wherein, cover plate 162 covering substrate 161, both are fixed in the through hole 152a of installing plate 152, electromagnetic valve 163 is installed on cover plate 162, refrigerating plant 164 is arranged on the base plate 151 of support 15 and is close to substrate 161, and heater 165 is folded between substrate 161 and cover plate 162 and is close to substrate 161.
Fig. 4 is the perspective view of cover plate, and cover plate 162 has rectangular body 1621, and this rectangular body 1621 is suitable for being contained in the through hole 152 of installing plate 152 carrying out installing fixes. Offering groove 1622 in this rectangular body 1621, the test section of substrate 161 is exposed from groove 1622, thus being exposed under the irradiation of synchrotron radiation light.
Fig. 5 is the top view of substrate, and on substrate 161, processing has microchannel, can be divided into constant temperature liquid storage district, Temperature jump district and synchrotron radiation X-ray test section. specifically, substrate 161 has the first relatively independent microchannel 1611, second microchannel 1612 and the 3rd microchannel 1613, wherein, the arrival end 1611a of the first microchannel 1611 is connected by conduit with syringe pump 13, the port of export 1611b of the first microchannel 1611 is connected with electromagnetic valve 163, the arrival end 1612a of the second microchannel 1612 is connected by conduit with the first container 141 of fluid supply 14, the port of export 1612b of the second microchannel 1612 is connected with electromagnetic valve 163, the arrival end 1613a of the 3rd microchannel 1613 is connected with electromagnetic valve 163, thus, electromagnetic valve 163 is used to connection the first microchannel 1611, second microchannel 1612 and the 3rd microchannel 1613. when the first microchannel 1611 is connected by electromagnetic valve 163 with the 3rd microchannel 1613, both collectively constitute constant temperature liquid storage microchannel. the downstream of the 3rd microchannel 1613 is sequentially connected with the 4th microchannel 1614 and the 5th microchannel 1615, after the outlet of the 3rd microchannel 1613 is divided into two-way, width increases, and the degree of depth reduces, extending and form the 4th microchannel 1614, the 4th microchannel 1614 is rectangle-trapezoidal microchannel in the present embodiment, it is behind a road that the outlet of the 4th microchannel 1614 is converged again, and width reduces, the degree of depth increases and forms the 5th microchannel 1615, and the port of export 1615b of the 5th microchannel 1615 is connected by conduit with the second container 142 of fluid supply 14. computer 11 is connected with syringe pump 13, electromagnetic valve 163 communication, controls the duty of syringe pump and electromagnetic valve thereby through computer 11, thus the flowing velocity of the fluid changed in microchannel and direction.
In conjunction with Fig. 3, refrigerating plant 164 and substrate 161 are in close contact and directly cool down the 1611, second microchannel, the first microchannel 1612 and the 3rd microchannel 1613. Heater 165 is in close contact with substrate 161 and heats the 4th microchannel 1614 equally, so that the 4th microchannel 1614 becomes Temperature jump district.5th microchannel 1615 is sealed by kapton film, forms synchrotron radiation X-ray test section. In the present embodiment, the position of the end of being close to the exit of the 5th microchannel 1615 is formed with waste liquid buffer memory microchannel 1616. In the present embodiment, the cover plate 162 corresponding to test position 1611c in the first microchannel 1611 leaves rectangular channel, the miniature warming probe (not shown) being connected with computer 11 communication is arranged in this rectangular channel of cover plate 162, thus by the temperature Real-time Feedback in constant temperature liquid storage microchannel to refrigerating plant 164, it is achieved the accurate temperature in constant temperature liquid storage microchannel controls. In the present embodiment, this electromagnetic valve 163 is two-bit triplet electromagnetic valve. In the present embodiment, the micro-thermode that adds of metal that this heater 165 is external, it is the Pt electrode of sputtering patterning on the glass substrate, thus the horizontal span meeting the thermal treatment zone is only the requirement of 0.5-1mm.
The concrete structure of micro flow chip system is risen in conjunction with above-mentioned fast temperature, the workflow of the present invention described further below: by chip 16 fixed support 15, then support 15 is fixed on two-dimension adjustment platform 12, horizontal level adjustment module 121 and upright position adjustment module 122 is regulated, by a certain position alignment synchrotron radiation hot spot of the 5th microchannel 1615 of chip 16 by computer 11. Sampling volume and the sample introduction speed of syringe pump 13 are set by computer 11 subsequently, first valve port 132 is set to sample introduction valve port, the duty of electromagnetic valve 163 is set by computer 11 simultaneously, make the port of export 1611b of the first microchannel 1611 and the port of export 1612b connection of the second microchannel 1612, after starting sample introduction operation, sample solution is drawn into the first microchannel 1611 by syringe pump 13 from the first container 141. Open refrigerating plant 164, in miniature warming probe display first microchannel 1611 at the test position 1611c place of the first microchannel 1611, liquid constant is after design temperature, the duty of electromagnetic valve 163 is set by computer 11, make the arrival end 1613a connection of port of export 1611b and the three microchannel 1613 of the first microchannel 1611, and control syringe pump 13 liquid in first microchannel 1611 is quickly propelled to the 4th microchannel 1614 via the 3rd microchannel 1613. Meanwhile, the control circuit of heater 165 connected by computer 11, and the sample solution of flowing of extending in the 4th microchannel 1614, with heater 165, comparatively strong heat convection occurs, and solution temperature rises to rapidly. Sample solution after temperature-jump continues to flow into the 5th microchannel 1615, opens beam shutter, carries out synchrotron radiation X-ray measurement.
After being measured, arranging sampling volume and the sample introduction speed of syringe pump 13, and the 3rd valve port 134 is set to sample introduction valve port, after starting sample introduction operation, cleanout fluid is drawn in syringe pump 13 by syringe pump 13 from the 3rd container 143. Setting the first valve port 132 is injection port, the duty of electromagnetic valve 163 is set by computer 11, make the arrival end 1613a connection of port of export 1611b and the three microchannel 1613 of the first microchannel 1611, after starting stock layout operation, the cleanout fluid that it is internal is advanced in micro flow chip passage by syringe pump 13, cleanout fluid, via behind the first microchannel 1614, microchannel the 1613, the 4th, microchannel the 1611, the 3rd and the 5th microchannel 1615, is drained into second container 142. It addition, when the second valve port 133 of syringe pump 13 is set to sample introduction valve port, same to aforesaid operations, air can be passed in micro flow chip passage, it is achieved microchannel functions/drying.
Above-described, it is only presently preferred embodiments of the present invention, is not limited to the scope of the present invention, the above embodiment of the present invention can also make a variety of changes.Namely every claims according to the present patent application and description are made change simple, equivalent and modification, fall within the claims of patent of the present invention. The not detailed description of the present invention be routine techniques content.
Claims (10)
1. a fast temperature rises to micro flow chip system, it is characterised in that including:
Chip, this chip includes substrate, cover plate, electromagnetic valve, refrigerating plant and heater, substrate has microchannel, this microchannel sequentially forms constant temperature liquid storage district, Temperature jump district and test section, cover plate covers and seal substrate, electromagnetic valve connection microchannel, and refrigerating plant is close to the constant temperature liquid storage district of substrate and is arranged, heater is close to the Temperature jump district of substrate and is arranged, and test section is exposed under the irradiation of synchrotron radiation light and tests;
For installing the support fixing this chip;
For regulating the two-dimension adjustment platform of the position of support, this two-dimension adjustment platform includes horizontal level adjustment module and upright position adjustment module; And
Syringe pump, this syringe pump has body and the first valve port being connected with this body, and described valve port connects with the microchannel of chip.
2. fast temperature according to claim 1 rises to micro flow chip system, it is characterized in that, microchannel includes relatively independent the first microchannel, the second microchannel and the 3rd microchannel, wherein, the arrival end of the first microchannel is connected with syringe pump, and the port of export of the first microchannel is connected with electromagnetic valve, and the arrival end of the second microchannel is connected with sample liquids, the port of export of the second microchannel is connected with electromagnetic valve, and the arrival end of the 3rd microchannel is connected with electromagnetic valve.
3. fast temperature according to claim 2 rises to micro flow chip system, it is characterized in that, microchannel also includes the 4th microchannel and the 5th microchannel that are set in turn in the 3rd downstream, microchannel, the 4th formation temperature saltus step district, microchannel, the 5th formation test section, microchannel.
4. fast temperature according to claim 3 rises to micro flow chip system, it is characterised in that the 4th microchannel is that width first disperses the microchannel restrained afterwards.
5. fast temperature according to claim 3 rises to micro flow chip system, it is characterised in that with first, second, third and the 5th compared with microchannel, and the width of the 4th microchannel is relatively big and the degree of depth is less.
6. fast temperature according to claim 2 rises to micro flow chip system, it is characterised in that this chip also includes the warming probe at the test position place being arranged at the first microchannel.
7. fast temperature according to claim 1 rises to micro flow chip system, it is characterised in that this syringe pump also includes the second valve port with air communication.
8. fast temperature according to claim 1 rises to micro flow chip system, it is characterised in that this syringe pump also includes the 3rd valve port connected with cleanout fluid.
9. fast temperature according to claim 1 rises to micro flow chip system, it is characterised in that this support includes base plate and is perpendicular to the installing plate of this base plate, and this installing plate has through hole, and chip is fixed in this through hole.
10. fast temperature according to claim 1 rises to micro flow chip system, it is characterised in that this heater is the Pt electrode of sputtering patterning on the glass substrate.
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| CN108614600B (en) * | 2018-07-13 | 2024-04-30 | 中国科学院天津工业生物技术研究所 | High-precision chip reaction system and method |
| CN109374908A (en) * | 2018-11-23 | 2019-02-22 | 中国科学院上海高等研究院 | A kind of vacuum automatic sample device and vacuum specimen chamber suitable for solution high flux screening |
| CN109374908B (en) * | 2018-11-23 | 2024-01-26 | 中国科学院上海高等研究院 | Vacuum automatic sample device and vacuum sample chamber suitable for high-throughput screening of solution |
| CN113670919A (en) * | 2021-07-30 | 2021-11-19 | 中国计量大学 | Micro-bubble visualization device and visualization method for micro-fluidic chip in temperature changing process |
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