CN111111799A - Method for hydrophilic modification of surface of microfluidic chip - Google Patents
Method for hydrophilic modification of surface of microfluidic chip Download PDFInfo
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- CN111111799A CN111111799A CN201911366649.4A CN201911366649A CN111111799A CN 111111799 A CN111111799 A CN 111111799A CN 201911366649 A CN201911366649 A CN 201911366649A CN 111111799 A CN111111799 A CN 111111799A
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Abstract
The invention provides a method for hydrophilic modification of the surface of a microfluidic chip, which comprises the following steps: (1) connecting the plasma treatment instrument with a plasma vaporizer; (2) pouring the hydrophilic agent into a charging cavity of a plasma vaporizer, setting the heating temperature of the hydrophilic agent to be 35-80 ℃, the heat preservation temperature of a wire tube to be 40-85 ℃, starting a power supply, preheating for 2-8 min, and waiting for grafting; (3) placing a micro-fluidic chip to be modified in a processing cavity of a plasma processing instrument, opening a valve of a plasma vaporizer, introducing a hydrophilic agent into the plasma processing instrument, vacuumizing to 10-80 pa, wherein the gas flow of the hydrophilic agent is 1.5-6L/min, the discharge power is 45-150W, and the plasma polymerization time is 0.5-5 min; (4) and after the reaction is finished, introducing air into the treatment cavity. The micro-fluidic chip prepared by the method has long hydrophilic retention time, good coating adhesion and no specific adsorption on target detection molecules.
Description
Technical Field
The invention belongs to the technical field of microfluidic chips, and particularly relates to a method for hydrophilic modification of the surface of a microfluidic chip.
Background
The microfluidic chip technology is a technology for manufacturing a microchannel network structure and other functional units on a very small (several square centimeters) chip by adopting a micro-machining technology, integrating basic operation units related to the fields of biology, chemistry and the like, such as sample preparation, biological and chemical reactions, separation, detection and the like, or basically integrating the basic operation units on an operation platform as small as possible, so as to complete different biological or chemical reaction processes and analyze products of the biological or chemical reaction processes.
High molecular polymer micro-fluidic chips represented by polymethyl methacrylate (PMMA) are widely used in the field of bioanalysis due to their advantages of high light transmittance, low cost, mass production, etc. However, most of microfluidic chips based on high molecular polymers have strong hydrophobicity, which not only seriously affects the entry of analytes into the channels and the mobility of analytes in the channels, but also causes inaccurate analysis due to the adsorption of biomolecules, thereby greatly limiting the range of analytes and reducing the practicability of the microfluidic chips.
In order to solve the problem of strong hydrophobicity of the surface of the microfluidic chip, appropriate hydrophilic modification is usually performed on the surface of the microfluidic chip so as to effectively control the size of electroosmotic flow (EOF), improve the hydrophilic performance of the surface of a micro-channel of the microfluidic chip, reduce the non-specific adsorption of target molecules on the surface of the micro-channel of the microfluidic chip, and obtain a more accurate analysis result.
The current hydrophilic modification technology of microfluidic chips can be roughly divided into the following categories: (1) directly adding hydrophilic materials into high polymer materials for preparing the micro-fluidic chip; (2) ultraviolet-O3Processing the microfluidic chip; (3) bombarding the chip by plasma; (4) a strong oxidation method; (5) surface chemical grafting, which can realize polymerization reaction on the surface of the chip by ultraviolet or other initiator induction, and grafting hydrophilic macromolecules; (6) a hydrophilic agent solution is applied. Although the prior methods for hydrophilic modification of the surface of the microfluidic chip are more, the following disadvantages exist: (1) the hydrophilic material is directly added into the high polymer material for preparing the microfluidic chip, and the hydrophilic material can generate chemical reaction when being melted at high temperature, so that the performance of the chip is influenced, and in addition, the hydrophilic material is not easy to be uniformly distributed in the material of the microfluidic chip and can also influence the uniformity of the microfluidic chip during ultrasonic welding; (2) ultraviolet-O3For handling microfluidic chipsThe method is convenient and quick, large-scale instruments and equipment are not needed, the cost is low, but the hydrophilicity of the chip can be receded along with time, the hydrophobicity is recovered, and in addition, the surface of the chip can be yellowed by ultraviolet irradiation; (3) the plasma bombardment method is convenient and rapid, but the hydrophilicity of the chip bombarded by the plasma can be removed along with time, and the hydrophobicity is recovered; (4) the strong oxidation method can obviously improve the hydrophilicity of the surface of the chip in a short time, but the method is not suitable for the chip with a microstructure or a microelectrode and other precise structures in a channel; (5) the surface chemical grafting method is usually carried out in an organic solvent, and the solvent may have influence on the chip material; (6) in the method for coating the hydrophilic agent solution, most of the hydrophilic agent is physically adsorbed on the surface of the chip, and the coating is very easy to fall off, so that the application of the microfluidic chip, such as biological modification, is not facilitated.
Therefore, a method for hydrophilic modification of the surface of the microfluidic chip with high efficiency, no damage and good adhesion is urgently needed to be developed.
Disclosure of Invention
Aiming at the problems that the surface modification method of the microfluidic chip in the prior art is short in hydrophilicity retention time, the chip is easy to damage in the modification process, the hydrophilic coating is easy to fall off and nonspecific adsorption is caused to target detection molecules, the invention provides the method for modifying the surface of the microfluidic chip, which has the advantages of high modification efficiency, long hydrophilicity retention time after modification, no damage to the chip, good coating adhesion and no specific adsorption to the target detection molecules.
In order to achieve the purpose, the invention provides the following technical scheme: a method for hydrophilic modification of the surface of a microfluidic chip comprises the following steps:
(1) connecting the plasma treatment instrument with a plasma vaporizer;
(2) pouring a hydrophilic agent into the charging cavity of the plasma vaporizer, setting the heating temperature of the hydrophilic agent to be 35-80 ℃, the heat preservation temperature of the wire tube to be 40-85 ℃, starting a power supply, preheating for 2-8 min, and waiting for grafting;
(3) placing a micro-fluidic chip to be modified in a processing cavity of a plasma processor, opening a valve of the plasma vaporizer, introducing a hydrophilic agent into the plasma processor, vacuumizing the plasma processor to 10-80 pa, wherein the gas flow of the hydrophilic agent is 1.5-6L/min, the discharge power is 45-150W, and the plasma polymerization time is 0.5-5 min;
(4) and after the reaction is finished, introducing air into the treatment cavity for 3min, and taking out the chip.
Further, the hydrophilic agent is one or two of polyacrylic acid, polyvinyl alcohol, polyvinylpyrrolidone, polyethyleneimine, waterborne polyurethane, acrylic acid, tetrabutyl titanate, butyl orthosilicate, hydroxyethyl methacrylate and dopamine.
Further, the plasma vaporizer comprises a plurality of charging chambers, and different hydrophilic agents are filled in the plurality of charging chambers. Because the required vaporization temperature of every kind of hydrophilic agent is different, set up a plurality of charging chambers and can guarantee that hydrophilic agent fully vaporizes, every charging chamber all has the spool of connecting the processing chamber, consequently can guarantee that a plurality of charging chambers advance the appearance simultaneously.
Further, in the step (3), the gas flow of the hydrophilic agent is 2.5L/min, the vacuum is pumped to 60pa, the discharge power is 45W, and the plasma polymerization time is 3.5 min.
Compared with the prior art, the invention has the following beneficial effects:
(1) compared with the traditional plasma treatment method, the hydrophilicity of the microfluidic chip provided by the invention is obviously improved after plasma polymerization treatment, mainly because the hydrophilic material is polymerized and grafted by the plasma, the hydrophilic material is combined with the surface of the microfluidic chip through chemical bonds, and the adhesive force between the coating and the chip is strong, so that the hydrophilicity of the microfluidic chip cannot deteriorate along with time, and a favorable basis is provided for the use of the microfluidic chip.
(2) The microfluidic chip prepared by the surface hydrophilic modification method has no damage to the chip in the modification process, the fluidity of the microfluidic chip is obviously improved, and the nonspecific adsorption of target detection molecules is reduced.
(3) The plasma polymerization modification method provided by the invention is convenient and rapid to operate, the difference between batches and within batches of the microfluidic chip is controllable, meanwhile, no waste is generated in the whole modification process, and the method is environment-friendly and suitable for industrial large-scale production.
Drawings
FIG. 1 is a comparison graph of the direct adsorption and chemical crosslinking effects of the modified microfluidic chip.
Detailed Description
The process of the present invention will be described in detail with reference to specific examples.
Method for hydrophilic modification of surface of microfluidic chip
Example 1
A method for hydrophilic modification of the surface of a microfluidic chip is characterized by comprising the following steps:
(1) communicating the plasma treatment instrument with a plasma gasification instrument;
(2) respectively pouring polyacrylic acid and n-butyl silicate into two charging cavities of a plasma vaporizer, screwing a charging cover, connecting a vaporizing tube between the plasma vaporizer and a plasma processor, and setting the heating temperature of the polyacrylic acid to be 35 ℃ and the heat preservation temperature of a wire tube to be 40 ℃; the heating temperature of the butyl orthosilicate is 65 ℃, and the temperature of the line pipe is 70 ℃; starting a power supply, preheating for 2min, and waiting for grafting;
(3) placing the micro-fluidic chip to be modified in a treatment cavity of a plasma treatment instrument, closing a cavity door, wherein the gas flow of polyacrylic acid is 1.5L/min, the gas flow of n-butyl silicate is 3L/min, vacuumizing to 40pa, the discharge power is 45W, and the plasma polymerization time is 3.5 min;
(4) and after the reaction is finished, introducing air into the treatment cavity for 3min, and taking out the chip.
Example 2
A method for hydrophilic modification of the surface of a microfluidic chip is characterized by comprising the following steps:
(1) communicating the plasma treatment instrument with a plasma gasification instrument;
(2) pouring polyethyleneimine into a charging cavity of a plasma vaporizer, screwing a charging cover, connecting a vaporizing tube between the plasma vaporizer and a plasma treatment instrument, and setting the heating temperature of the polyethyleneimine to be 80 ℃ and the heat preservation temperature of a wire tube to be 85 ℃; starting a power supply, preheating for 8min, and waiting for grafting;
(3) placing the micro-fluidic chip to be modified in a treatment cavity of a plasma treatment instrument, closing a cavity door, vacuumizing to 80pa with the gas flow of a hydrophilic agent being 6L/min, discharging power being 150W, and polymerizing plasma for 5 min;
(4) and after the reaction is finished, introducing air into the treatment chamber for 3min, and taking out the chip.
Example 3
A method for hydrophilic modification of the surface of a microfluidic chip is characterized by comprising the following steps:
(1) communicating the plasma treatment instrument with a plasma gasification instrument;
(2) respectively pouring acrylic acid and tetrabutyl titanate into two material loading cavities of a plasma vaporizer, screwing a material loading cover, connecting a vaporizing tube between the plasma vaporizer and a plasma processor, and simultaneously setting the heating temperature of the acrylic acid to be 35 ℃ and the heat preservation temperature of a wire tube to be 40 ℃; the heating temperature of tetrabutyl titanate is 55 ℃, and the temperature of the line pipe is 60 ℃; starting a power supply, preheating for 5min, and waiting for grafting;
(3) placing the micro-fluidic chip to be modified in a treatment cavity of a plasma treatment instrument, closing a cavity door, wherein the gas flow of acrylic acid is 3L/min, the gas flow of tetrabutyl titanate is 2.5L/min, vacuumizing to 10pa, the discharge power is 95W, and the plasma polymerization time is 30 s;
(4) and after the reaction is finished, introducing air into the treatment chamber for 3min, and taking out the chip.
Example 4
A method for hydrophilic modification of the surface of a microfluidic chip is characterized by comprising the following steps:
(1) communicating the plasma treatment instrument with a plasma gasification instrument;
(2) pouring dopamine into a charging cavity of a plasma vaporizer, screwing a charging cover, connecting a vaporizing tube between the plasma vaporizer and a plasma processor, setting the heating temperature of the dopamine to be 60 ℃, the heat preservation temperature of the tube to be 65 ℃, preheating for 6min, and waiting for grafting;
(3) placing the micro-fluidic chip to be modified in a treatment cavity of a plasma treatment instrument, closing a cavity door, vacuumizing to 60pa with the gas flow of a hydrophilic agent being 6.5L/min, discharging power being 150W, and polymerizing plasma for 2 min;
(4) and after the reaction is finished, introducing air into the treatment chamber for 3min, and taking out the chip.
Example 5
A method for hydrophilic modification of the surface of a microfluidic chip is characterized by comprising the following steps:
(1) communicating the plasma treatment instrument with a plasma gasification instrument;
(2) pouring the waterborne polyurethane into a charging cavity of a plasma vaporizer, screwing a charging cover, connecting a vaporizing tube between the plasma vaporizer and a plasma processor, setting the heating temperature of the waterborne polyurethane to be 50 ℃, the heat preservation temperature of a wire tube to be 55 ℃, starting a power supply, preheating for 4min, and waiting for grafting;
(3) placing the micro-fluidic chip to be modified in a treatment cavity of a plasma treatment instrument, closing a cavity door, vacuumizing to 40pa with the gas flow of a hydrophilic agent being 4.5L/min, discharging power being 120W, and polymerizing plasma for 1.5 min;
(4) and after the reaction is finished, introducing air into the treatment chamber for 3min, and taking out the chip.
Example 6
A method for hydrophilic modification of the surface of a microfluidic chip is characterized by comprising the following steps:
(1) communicating the plasma treatment instrument with a plasma gasification instrument;
(2) respectively pouring polyvinyl alcohol and polyethyleneimine into two charging cavities of a plasma vaporizer, screwing a charging cover, connecting a vaporizing tube between the plasma vaporizer and a plasma processor, and setting the heating temperature of the polyvinyl alcohol to be 60 ℃ and the heat preservation temperature of a wire tube to be 65 ℃; the heating temperature of the polyethyleneimine is 80 ℃, and the temperature of the line pipe is 85 ℃; starting a power supply, preheating for 6min, and waiting for grafting;
(3) placing the micro-fluidic chip to be modified in a treatment cavity of a plasma treatment instrument, closing a cavity door, wherein the gas flow of polyvinyl alcohol is 2L/min, the gas flow of polyethyleneimine is 3.5L/min, vacuumizing to 40pa, the discharge power is 120W, and the plasma polymerization time is 3 min;
(4) and after the reaction is finished, introducing air into the treatment chamber for 3min, and taking out the chip.
Example 7
A method for hydrophilic modification of the surface of a microfluidic chip is characterized by comprising the following steps:
(1) communicating the plasma treatment instrument with a plasma gasification instrument;
(2) respectively pouring polyvinylpyrrolidone and hydroxyethyl methacrylate into two charging cavities of a plasma vaporizer, screwing a charging cover, connecting a vaporizing tube between the plasma vaporizer and a plasma processor, and setting the heating temperature of the polyvinylpyrrolidone to be 50 ℃ and the heat preservation temperature of a wire tube to be 55 ℃; heating hydroxyethyl methacrylate at 35 deg.C, and keeping the temperature of the line pipe at 40 deg.C; starting a power supply, preheating for 3.5min, and waiting for grafting;
(3) placing the micro-fluidic chip to be modified in a treatment cavity of a plasma treatment instrument, closing a cavity door, wherein the gas flow of polyvinylpyrrolidone is 1.5L/min, the gas flow of hydroxyethyl methacrylate is 4L/min, vacuumizing to 60pa, the discharge power is 100W, and the plasma polymerization time is 2 min;
(4) and after the reaction is finished, introducing air into the treatment chamber for 3min, and taking out the chip.
Example 8
A method for hydrophilic modification of the surface of a microfluidic chip is characterized by comprising the following steps:
(1) communicating the plasma treatment instrument with a plasma gasification instrument;
(2) respectively pouring polyvinylpyrrolidone and tetrabutyl titanate into two charging cavities of a plasma vaporizer, screwing a charging cover, connecting a vaporizing tube between the plasma vaporizer and a plasma processor, and setting the heating temperature of the polyvinylpyrrolidone to be 50 ℃ and the heat preservation temperature of a wire tube to be 55 ℃; the heating temperature of tetrabutyl titanate is 55 ℃, and the temperature of the line pipe is 60 ℃; starting a power supply, preheating for 3min, and waiting for grafting;
(3) placing the micro-fluidic chip to be modified in a treatment cavity of a plasma treatment instrument, closing a cavity door, wherein the gas flow of polyvinylpyrrolidone is 1L/min, the gas flow of hydroxyethyl methacrylate is 3L/min, vacuumizing to 80pa, the discharge power is 110W, and the plasma polymerization time is 3 min;
(4) and after the reaction is finished, introducing air into the treatment chamber for 3min, and taking out the chip.
Secondly, verifying the effect of the surface hydrophilicity of the microfluidic chip obtained by adopting the modification method of the invention
2.1) determination of hydrophilic Angle of microfluidic chip
Determining the hydrophilic angle of the 5 micro-fluidic chips which are not processed to obtain the hydrophilic angle; then, the 5 micro-fluidic chips are subjected to hydrophilic modification treatment according to the surface hydrophilic modification method of the invention, and then the hydrophilic angle is tested again, 5 points are tested on each chip, and the results are shown in table 1:
table 1 test results of hydrophilic angle of microfluidic chip before and after surface hydrophilic modification treatment
As can be seen from table 1 above, the hydrophilic angle of the microfluidic chip prepared by the surface hydrophilic modification method of the present invention is significantly reduced, which indicates that the hydrophilicity of the microfluidic chip can be significantly improved by performing the surface hydrophilic modification on the microfluidic chip by the modification method of the present invention.
2.2) determination of adsorption stability of hydrophilic coating of microfluidic chip
After the 5 pieces of microfluidic chip with the modified surfaces in the 2.1) are subjected to ultrasonic treatment for 3min, the hydrophilic angle of the microfluidic chip is continuously tested, and the experimental results are shown in table 2:
table 2 hydrophilic angle test results of microfluidic chip after 3min of ultrasound
As can be seen from table 2 above, the hydrophilic coating of the microfluidic chip prepared by the present invention has good adsorption stability, and the hydrophilic angle does not greatly increase after 3min of ultrasound, which indirectly indicates that the coating does not fall off.
2.3) testing the hydrophilic stability of the microfluidic chip
Taking 5 tablet cores, carrying out hydrophilic modification treatment according to the surface hydrophilic modification method of the invention, testing the hydrophilic angle at intervals of time (1d, 10d, 30d, 90d, 180d and 360d), and testing 5 points for each tablet core, wherein the results are as follows:
table 3 hydrophilic angle test results of microfluidic chip with different retention times
As can be seen from table 4 above, the hydrophilic angle of the microfluidic chip prepared by the present invention does not increase with the increase of the storage time, which indicates that the surface hydrophilic modification of the microfluidic chip by the modification method of the present invention can obtain permanent hydrophilicity.
2.4) determination of the flowability of microfluidic chips
The microfluidic chip with the tested hydrophilic angle in 2.1) is added with a blood filtering film for welding and sealing, and then added with anticoagulated whole blood for testing the fluidity of the microfluidic chip, and the results are shown in table 3:
table 4 flow time of anticoagulated whole blood through microfluidic chip
| Chip numbering | 1 | 2 | 3 | 4 | 5 |
| Flow time/s | 396 | 422 | 405 | 385 | 361 |
As can be seen from the above Table 4, in the microfluidic chip subjected to the surface hydrophilic modification treatment of the present invention, the flow time of the anticoagulated whole blood through the micro channel is 6-8min, while the unmodified microfluidic chip and the anticoagulated whole blood cannot flow, and after standing for 1h, no blood sample passes through the microfluidic chip. This shows that the fluidity of the microfluidic chip is significantly improved after the surface modification treatment of the present invention.
2.5) adsorption assay of micro-fluidic chip biomolecules
The method comprises the steps of adding two sites on a micro-channel of a micro-fluidic chip by using a spotting instrument, directly adding a mouse antibody at the site 1 without using a cross-linking agent, cross-linking the mouse antibody at the site 2 by using a biological cross-linking agent, adding a layer of 0.5% BSA (bovine serum albumin) in a mixed region, then adding a fluorescent secondary antibody, adding a blood filter membrane and welding a sealing sheet, and adding anticoagulated whole blood to test the adsorption property of the micro-fluidic chip on biomolecules, wherein the result is shown in figure 1. As can be seen, only site 2 where the biological crosslinker was used was able to capture the fluorescent secondary antibody. This is mainly because site 1 is an antibody added through a physical adsorption site, site 2 is an antibody crosslinked by a chemical crosslinking agent, and when the antibody is directly added on a chip in a weak nonspecific state, the antibody cannot be fixed on the chip, so that when the fluorescent secondary antibody is used for verification, site 1 (physical adsorption) cannot capture the fluorescent secondary antibody because no antibody is fixed, and site 2 can capture the fluorescent secondary antibody because the antibody is fixed by the crosslinking agent. The micro-fluidic chip subjected to surface hydrophilic modification treatment can obviously reduce the adsorption to biological molecules and has no specific adsorption to target detection molecules.
Finally, the above embodiments are only for illustrating the technical solutions of the present invention and not for limiting, although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions may be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all of them should be covered in the claims of the present invention.
Claims (4)
1. A method for hydrophilic modification of the surface of a microfluidic chip is characterized by comprising the following steps:
(1) connecting the plasma treatment instrument with a plasma vaporizer;
(2) pouring a hydrophilic agent into the charging cavity of the plasma vaporizer, setting the heating temperature of the hydrophilic agent to be 35-80 ℃, the heat preservation temperature of the wire tube to be 40-85 ℃, starting a power supply, preheating for 2-8 min, and waiting for grafting;
(3) placing a micro-fluidic chip to be modified in a processing cavity of a plasma processor, opening a valve of the plasma vaporizer, introducing a hydrophilic agent into the plasma processor, vacuumizing the plasma processor to 10-80 pa, wherein the gas flow of the hydrophilic agent is 1.5-6L/min, the discharge power is 45-150W, and the plasma polymerization time is 0.5-5 min;
(4) and after the reaction is finished, introducing air into the treatment cavity for 3min, and taking out the chip.
2. The method for hydrophilic modification of the surface of a microfluidic chip according to claim 1, wherein the hydrophilic agent is one or two of polyacrylic acid, polyvinyl alcohol, polyvinylpyrrolidone, polyethyleneimine, waterborne polyurethane, acrylic acid, tetrabutyl titanate, tetrabutyl orthosilicate, hydroxyethyl methacrylate and dopamine.
3. The method for hydrophilic modification of the surface of the microfluidic chip according to claim 1, wherein the plasma vaporizer comprises a plurality of charging chambers, and different hydrophilic agents are filled in the plurality of charging chambers.
4. The method for hydrophilic modification of the surface of the microfluidic chip according to claim 1, wherein the gas flow rate of the hydrophilic agent in the step (3) is 2.5L/min, the vacuum is pumped to 60pa, the discharge power is 45W, and the plasma polymerization time is 3.5 min.
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| CN108080045A (en) * | 2017-12-27 | 2018-05-29 | 北京百奥芯科技有限公司 | A kind of hydrophilically modified method of fluid channel inside micro-fluidic chip |
| CN108579828A (en) * | 2018-04-18 | 2018-09-28 | 清华大学天津高端装备研究院 | A kind of Surface modification of microfluidic chip method that flow velocity is controllable |
| CN110433881A (en) * | 2019-09-02 | 2019-11-12 | 丹娜（天津）生物科技有限公司 | A kind of hydrophilic modification method of micro-fluidic chip microchannel material |
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