CN107308996B - Method for maintaining negative pressure state of micro-fluidic chip for long time - Google Patents
Method for maintaining negative pressure state of micro-fluidic chip for long time Download PDFInfo
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- CN107308996B CN107308996B CN201710475437.4A CN201710475437A CN107308996B CN 107308996 B CN107308996 B CN 107308996B CN 201710475437 A CN201710475437 A CN 201710475437A CN 107308996 B CN107308996 B CN 107308996B
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- 229920006254 polymer film Polymers 0.000 claims abstract description 22
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- 239000004205 dimethyl polysiloxane Substances 0.000 claims description 24
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 claims description 24
- 238000000151 deposition Methods 0.000 claims description 17
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- 238000007740 vapor deposition Methods 0.000 claims description 10
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- -1 polydimethylsiloxane Polymers 0.000 claims description 3
- 238000005137 deposition process Methods 0.000 claims description 2
- 229920000052 poly(p-xylylene) Polymers 0.000 claims description 2
- 239000012808 vapor phase Substances 0.000 claims description 2
- 238000002347 injection Methods 0.000 abstract description 14
- 239000007924 injection Substances 0.000 abstract description 14
- 238000001514 detection method Methods 0.000 abstract description 9
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- 238000007789 sealing Methods 0.000 abstract description 4
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- 238000003860 storage Methods 0.000 abstract description 2
- VRBFTYUMFJWSJY-UHFFFAOYSA-N 28804-46-8 Chemical compound ClC1CC(C=C2)=CC=C2C(Cl)CC2=CC=C1C=C2 VRBFTYUMFJWSJY-UHFFFAOYSA-N 0.000 description 24
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- 108020004463 18S ribosomal RNA Proteins 0.000 description 1
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- DEGAKNSWVGKMLS-UHFFFAOYSA-N calcein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(CN(CC(O)=O)CC(O)=O)=C(O)C=C1OC1=C2C=C(CN(CC(O)=O)CC(=O)O)C(O)=C1 DEGAKNSWVGKMLS-UHFFFAOYSA-N 0.000 description 1
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- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502707—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the manufacture of the container or its components
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Abstract
The invention provides a method for maintaining a negative pressure state of a microfluidic chip for a long time. The polymer film has the characteristics of compactness, low permeability to gas molecules, no color, transparency and no fluorescence, so that the long-time storage of the negative pressure driving capability of the micro-fluidic chip is realized, the negative pressure driving micro-fluidic chip can be widely applied, the micro-fluidic chip without an external power source can be more favorably used for negative pressure driving sample injection, and the polymer film is applied to remote areas and on-site timely detection. During the sample introduction period of the chip, the deposited polymer film can still continuously protect the periphery of the chip from being in a sealing state, so that gas molecules are prevented from entering the chip from the periphery of the chip, the sample introduction effect is better, and the evaporation of reagents is prevented.
Description
Technical Field
The invention belongs to detection methods in multiple fields of life science, clinical diagnosis, chemical analysis and the like, and relates to a method for maintaining a negative pressure state of a microfluidic chip for a long time.
Background
The micro-fluidic chip technology is widely applied to various fields such as biology, chemistry, medicine and the like. Compared with the traditional laboratory detection and analysis method, the microfluidic chip technology has the advantages of small sample amount, simple operation, high flux, high accuracy, no cover opening, low pollution and the like, and is a great direction for the development of future detection technology.
In practical application, the micro-fluidic chip mostly needs an external power source to provide power to drive the sample to flow in the chip, the external power source is usually large in size and complex in structure, development and use costs are increased, the micro-fluidic chip is not easy to carry, and inconvenience is brought to on-site timely detection. A negative pressure driving sample injection chip without an external power source is provided, and the problem is well solved. The negative pressure driven micro-fluidic chip is made of Polydimethylsiloxane (PDMS), and the chip is vacuumized in advance by utilizing the characteristic that the porous gas storing body of the PDMS material can store gas, so that the internal pressure of the chip is lower than the external atmospheric pressure. Therefore, when the sample reagent is added at the inlet of the chip, the sample solution can be sucked into the chip due to the pressure difference between the inside and the outside of the chip, and an external power source is not needed. However, such a chip also has some disadvantages during use: firstly, the chip needs to be used immediately after each vacuumizing, which brings inconvenience to the remote area and the site instant detection; secondly, when the chip is used, gas molecules in the air can be sucked into the chip again from the periphery of the chip, and the sample introduction effect of the chip is influenced.
Disclosure of Invention
The invention aims to provide a method for maintaining the negative pressure state of a microfluidic chip for a long time, which is characterized in that a polymer film is deposited on the surface of the microfluidic chip, and the polymer film has the characteristics of compactness, low permeability to gas molecules, no color, transparency and no fluorescence, so that the long-time storage of the negative pressure driving capability of the microfluidic chip is realized, and the negative pressure driving microfluidic chip can be widely applied; during the sample injection period of the chip, the deposited polymer film can still continuously protect the periphery of the chip in a sealed state, and gas molecules are prevented from entering the chip from the periphery of the chip.
The invention provides a method for maintaining a negative pressure state of a microfluidic chip for a long time. The polymer film has the characteristics of compactness, low permeability to gas molecules, colorless transparency and no fluorescence.
The material of the micro-fluidic chip is Polydimethylsiloxane (PDMS).
The micro-fluidic chip obtains negative pressure driving force in a vacuumizing mode.
The polymer film deposited on the surface of the microfluidic chip is poly (p-xylylene chloride) (Parylene C).
The thickness of the microfluidic chip is adjustable, and is preferably 3-7 mm.
The thickness of the Parylene C film deposited on the surface of the microfluidic chip is adjustable, and is preferably 10-15 μm.
The polymer film is deposited on the surface of the microfluidic chip by using a vacuum vapor deposition method.
And vacuumizing the microfluidic chip and depositing the polymer film simultaneously.
Before the Parylene C film is deposited on the microfluidic chip in a vacuum vapor phase mode, a layer of transparent adhesive tape needs to be attached to the upper surface of the microfluidic chip.
The pressure of a deposition chamber of the vacuum vapor deposition equipment is adjustable, and the pressure is kept fixed during the deposition process.
The thickness of the transparent adhesive tape is not less than 0.05mm, namely more than or equal to 0.05 mm.
The invention has the advantages that: (1) the negative pressure power of the micro-fluidic chip can be kept for a long time, the time can reach 30 days or longer, the micro-fluidic chip without an external power source is more beneficial to sample injection by negative pressure driving, and the micro-fluidic chip is applied to timely detection in remote areas and sites; (2) the polymer film is not required to be removed when the sample is injected into the chip, the periphery of the chip can be kept in a sealed state by the film in the sample injection process of the chip, the sample injection effect is better, and the reagent can be prevented from evaporating to a certain extent; (3) the polymer film is colorless and transparent, has no fluorescence, and cannot influence the result detection of biochemical reaction in the chip.
Drawings
FIG. 1 is a flow chart of the method of the present invention.
Fig. 2 is a schematic diagram of example 2, wherein 1 is a microfluidic chip, 2 is an adhesive tape on the upper surface, 3 is a deposited Parylene C film, 4 is a sample inlet, and 5 is a nut-up position.
FIG. 3 is a graph showing the results of example 2.
FIG. 4 is a graph showing the results of example 3.
FIG. 5 is a schematic view of example 4, wherein 10 is a negative pressure driven pump module, which is composed of an upper surface adhesive tape 2, a deposited Parylene C film 3, a negative pressure driven PDMS module 5, and a lower surface adhesive tape 6; 7 is a micro-fluidic chip, 8 is a sample inlet, 9 is a sample outlet, and 11 is a negative pressure driving pump module with the lower surface adhesive tape removed.
Detailed Description
The invention is further illustrated with reference to the following figures and examples.
Example 1 Process flow
Referring to fig. 1, an implementation flow of a method for maintaining a negative pressure state of a microfluidic chip for a long time. The implementation object is a microfluidic chip, and the specific method is to stick a layer of transparent adhesive tape on the upper surface of the manufactured microfluidic chip and then deposit a layer of polymer film on the surface of the microfluidic chip stuck with the transparent adhesive tape.
Referring to fig. 2, the left side of the drawing is a schematic cross-sectional view, a layer of transparent adhesive tape 2 is adhered on the upper surface of the microfluidic chip 1, a layer of Parylene C film 3 is deposited on the surface of the microfluidic chip adhered with the transparent adhesive tape 2, the right side of the drawing is a schematic upper surface of the chip, wherein the sample injection ports 4 are arranged on two sides, meanwhile, nuts 5 are arranged in the middle positions of the two sides as support pads to lift the microfluidic chip 1, and the nuts 5 can be used in standard sizes of M6 to M10.
The material of the micro-fluidic chip is a polymer material with air permeability, preferably PDMS. The size of the chip can be adjusted as required. The thickness of the chip can be adjusted according to requirements, and is preferably 3-7 mm.
Before the polymer film is deposited on the microfluidic chip, a layer of transparent adhesive tape is adhered to the upper surface of the microfluidic chip, so that the problems of upper surface blurring, sample outlet blockage, insufficiently tight chip sealing and the like caused by the fact that the polymer film is directly deposited on the upper surface of the chip in the manufacturing process are solved. The thickness of the transparent adhesive tape is not less than 0.05mm, and the transparent adhesive tape is prevented from deforming and being incapable of being restored during vacuum pumping.
The deposited polymer film material has dense, low permeability to gas molecules, colorless, transparent, non-fluorescent properties, preferably Parylene C. The thickness of the polymer film can be adjusted as desired, preferably 10 to 15 μm.
The process of thin film deposition requires that a vacuum be maintained, and a vacuum vapor deposition method is preferred.
The equipment requirements for vacuum vapor deposition are that the pressure of the deposition chamber can be adjusted and kept constant while the deposition is in progress. The pressure may be in the range of 10 to 40mTorr, preferably 20 mTorr. When the microfluidic chip is placed in a vacuum vapor deposition system, the bottom surface needs to be suspended, and the Parylene C film can be uniformly deposited.
Example 2 method for maintaining a Long-time negative pressure on a self-priming liquid-separation digital polymerase chain reaction (digital PCR) chip
Referring to fig. 2, a method for maintaining a negative pressure state of a microfluidic chip for a long time is used for a self-priming liquid-separating digital PCR chip. The chip consists of a microfluidic chip 1, an adhesive tape 2 on the upper surface of the chip and a Parylene C film 3 deposited on the surface of the chip.
The micro-fluidic chip 1 adopts PDMS as a material, adopts a multilayer soft lithography technology to manufacture a mold with a micro-channel and a micro-reaction chamber on the surface of a silicon wafer by using optical cement, a PDMS thin layer with the thickness of 0.5mm is firstly spin-coated on the mold, and the proportion of PDMS is monomer: curing agent 5: 1. Baking at 85 deg.C for 2min, coating an anti-evaporation layer, and baking at 85 deg.C for 1 min. Then, a layer of PDMS with the thickness of 5mm is poured on the evaporation prevention layer, and the proportion of the PDMS is as follows: curing agent 10:1, and baking at 85 deg.C for 40 min. The PDMS monolith was removed from the mold and an inlet 4 was punched out using a 1mm diameter punch. And (3) sealing the cover glass with PDMS after the cover glass is treated by plasma, and then baking the cover glass on a hot plate at 85 ℃ for 4 hours to finish the processing of the chip.
A transparent adhesive tape 2 with the thickness of 0.07mm is stuck on the upper surface of the manufactured microfluidic chip 1, and a scraper is used for removing air bubbles between the transparent adhesive tape and the upper surface of the chip. Trimming the adhesive tape around the chip after the back edge is adhered.
The Parylene C film 3 deposited on the surface of the chip is deposited by using a vacuum vapor deposition system. And spraying a release agent in a deposition cavity of the vacuum vapor deposition system, and uniformly coating the release agent. Two nuts are used for each chip to be cushioned at position 5, so that the bottom surface is kept suspended, and the Parylene C film can be deposited on the bottom surface of the chip. 16.66g of Parylene C raw material was weighed, placed in a tin paper roll, and inserted into a raw material bin. 16.66g of Parylene C starting material can yield a film 10 μm thick. The cold trap is opened first, then the vacuum pump is opened, and the whole deposition system is vacuumized until the pressure of the deposition chamber reaches below 10mTorr, namely <10 mTorr. And opening heating knobs of the cracking cavity and the evaporation cavity, and starting the deposition of the Parylene C film when the temperature of the cracking cavity rises to 690 ℃, the temperature of the evaporation cavity reaches 135 ℃. At this time, the pressure in the deposition chamber was 20Pa, and 20Pa was maintained during the deposition.
After the raw materials in the raw material bin are completely evaporated and cracked, the pressure in the deposition cavity is gradually reduced, the temperatures in the cracking cavity, the evaporation cavity and the deposition cavity start to fall back, when the temperature of the cracking cavity is reduced to be lower than 350 ℃, the system is closed, and when the pressure in the deposition cavity reaches the atmospheric pressure, the chip is taken out. At this time, a layer of uniform, dense and transparent Parylene C film with a thickness of 10 μm has been deposited on the surface of the chip.
The chips deposited with the Parylene C coating were stored at atmospheric pressure at room temperature for 30 days. The injection port 4 is pricked by using a syringe needle, the human 18S ribosomal RNA plasmid and the matched reaction reagent of PCR are added, then the oil phase liquid which is incompatible with the reaction reagent is added, the injection port 4 is closed, and the digital PCR reaction is carried out in the chip, and the result is shown in figure 3.
Example 3 investigation of Parylene C film thickness on negative pressure holding time of microfluidic chip
In this example, the apparatus of example 2 was used to inject samples, and 8.33g, 16.66g and 24.99g of raw materials were used to deposit Parylene C films, respectively, to obtain 5 μm, 10 μm and 15 μm thick Parylene C films.
On the first day, the second day, and the thirty-third day, the microfluidic chips coated with Parylene C films with different thicknesses are taken respectively, the injection port 4 is pricked open by using a syringe needle, the calcein fluorescent dye is added, then the oil phase liquid is added for sealing, and the injection time is counted.
The microfluidic chip manufactured by the sample injection device in example 2, on which the Parylene C film is not deposited, is placed for 18 hours after being vacuumized, and thus complete sample injection of the fluorescent dye cannot be completed.
The microfluidic chip deposited with the Parylene C film can be kept for 30 days and still can be completely injected. The results are shown in FIG. 4.
EXAMPLE 4 method for maintaining Power of negative pressure driven Pump for a Long period of time
A method for maintaining the power of a negative pressure drive pump for a long time is used for sample injection analysis of a common micro-fluidic chip. The device consists of a negative pressure driving pump module 10 and a micro-fluidic chip 7. The negative pressure driving pump module 10 consists of an adhesive tape 2 on the upper surface, a deposited Parylene C film 3, a negative pressure driving PDMS module 5 and an adhesive tape 6 on the lower surface; the micro-fluidic chip 7 comprises a sample inlet 8 and a sample outlet 9; 11 is a negative pressure driven pump module with the lower surface adhesive tape removed.
The negative pressure driving pump module 10 adopts PDMS as a material, and is prepared by the following steps: the proportion of the curing agent is 10: 1. And pouring a PDMS module with the height of 5mm by using a clean silicon wafer as a bottom surface, and baking for 1h at 85 ℃ until the PDMS module is completely cured. Cut into a rectangular parallelepiped having a width equal to that of the chip 7 and a length of about 20 mm. The PDMS block 5 is driven by negative pressure.
A transparent adhesive tape 2 with the thickness of 0.07mm is pasted on the upper surface of the prepared negative pressure drive PDMS module 5, and a transparent adhesive tape 6 with the thickness of 0.07mm is pasted on the lower surface of the prepared negative pressure drive PDMS module. The air bubbles between the scotch tape and the upper surface of the chip were removed using a squeegee.
The Parylene C film 3 was deposited on the surface of the chip using the vacuum vapor deposition method of example 2.
And adding a reagent to be detected into the sample inlet 8, and removing the lower surface adhesive tape 6 of the negative pressure driving pump module 10 to expose the bottom surface of the PDMS, thereby obtaining the negative pressure driving module 11 with the lower surface adhesive tape removed. The side of the negative pressure driving pump module 11, from which the lower surface tape is removed, exposed out of the bottom surface of the PDMS is attached to the sample outlet 9. The negative pressure power provided by the negative pressure driving pump module can suck the reagent to be detected in the sample inlet 8 into the chip, and finally flows to the sample outlet 9 to finish detection. The results are shown in FIG. 5.
Claims (4)
1. A method for maintaining a negative pressure state of a microfluidic chip for a long time is characterized in that a layer of transparent adhesive tape (2) is pasted on the upper surface of the microfluidic chip (1), then a layer of polymer film (3) is deposited on the surface of the microfluidic chip pasted with the transparent adhesive tape (2), during deposition, the microfluidic chip (1) needs to be raised on the periphery, most of the central area of the lower surface of the chip is kept suspended, and the microfluidic chip (1) obtains a negative pressure driving force in a vacuumizing mode; the polymer film (3) is deposited on the surface of the microfluidic chip (1) by using a vacuum vapor deposition method; vacuumizing the microfluidic chip (1) and depositing the polymer film (3) are completed simultaneously; before the polymer film (3) is deposited in a vacuum vapor phase manner on the microfluidic chip (1), a layer of transparent adhesive tape (2) needs to be adhered to the upper surface of the microfluidic chip, and the thickness of the transparent adhesive tape (2) is more than or equal to 0.05 mm.
2. The method for maintaining the negative pressure state of the microfluidic chip for a long time according to claim 1, wherein the material of the microfluidic chip (1) is polydimethylsiloxane with the thickness of 3-7 mm.
3. The method for maintaining the negative pressure state of the microfluidic chip for a long time as claimed in claim 1, wherein the polymer film (3) is made of poly-p-xylylene monochloride and has a surface deposition thickness of 10-15 μm.
4. The method for maintaining the negative pressure state of the microfluidic chip for a long time according to claim 1, wherein the pressure of the deposition chamber during the vacuum vapor deposition is kept constant during the deposition process.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102395830A (en) * | 2009-03-13 | 2012-03-28 | 戈尔企业控股股份有限公司 | Moisture resistant coatings for polymeric enclosures |
| CN103071548A (en) * | 2012-04-05 | 2013-05-01 | 浙江大学 | Power source-free and valve-free type single molecule detection chip and applications thereof |
| CN106531646A (en) * | 2016-12-26 | 2017-03-22 | 中国科学院长春光学精密机械与物理研究所 | Method for packaging microfluidic chip |
| CN206215248U (en) * | 2016-11-07 | 2017-06-06 | 华南理工大学 | A kind of micro-fluidic chip of the microfluid spontaneous vasomotion of negative pressure guiding |
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| FR2848477B1 (en) * | 2002-12-17 | 2006-03-24 | Commissariat Energie Atomique | METHOD AND DEVICE FOR CONTAINING A LIQUID |
| KR100613398B1 (en) * | 2003-11-25 | 2006-08-17 | 한국과학기술연구원 | Element detecting system using cantilever, method for fabrication of the same system and method for detecting micro element using the same system |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102395830A (en) * | 2009-03-13 | 2012-03-28 | 戈尔企业控股股份有限公司 | Moisture resistant coatings for polymeric enclosures |
| CN103071548A (en) * | 2012-04-05 | 2013-05-01 | 浙江大学 | Power source-free and valve-free type single molecule detection chip and applications thereof |
| CN206215248U (en) * | 2016-11-07 | 2017-06-06 | 华南理工大学 | A kind of micro-fluidic chip of the microfluid spontaneous vasomotion of negative pressure guiding |
| CN106531646A (en) * | 2016-12-26 | 2017-03-22 | 中国科学院长春光学精密机械与物理研究所 | Method for packaging microfluidic chip |
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