CN115254209B - Preparation method of PDMS-PDA-MOFs micro-fluidic chip for single cell sequencing - Google Patents
Preparation method of PDMS-PDA-MOFs micro-fluidic chip for single cell sequencing Download PDFInfo
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Abstract
The invention relates to a preparation method of a PDMS-PDA-MOFs micro-fluidic chip for single cell sequencing, which is characterized in that an amino modified PDMS substrate is prepared; under alkaline condition, DA (dopamine) solution is subjected to self-polymerization on an amino modified PDMS substrate to obtain a PDMS substrate modified by PDA (polydopamine); then modifying MOFs on the surface of the modified polymer matrix to obtain a microsphere PDMS-PDA-MOFs elastomer with a microstructure coated inside and outside; and curing, attaching and baking the PDMS-PDA-MOFs elastomer in a mold to finally obtain the PDMS-PDA-MOFs microfluidic chip. Solves the problems of preparation and application of PDMS in the field of microfluidic chips.
Description
Technical Field
The invention relates to a preparation method of a PDMS-PDA-MOFs micro-fluidic chip, in particular to a preparation method of a PDMS-PDA-MOFs micro-fluidic chip for single cell sequencing.
Background
The heterogeneity of individual cells in a cell population plays an important role in the development and progression of disease, but most conventional genetic analysis methods now mask the differences in individual cells. Single cell sequencing can exhibit the inherent heterogeneity of single cells and reveal complex and rare cell populations. In recent years, different microfluidic technologies have emerged for single cell research, becoming the leading edge of the field.
Compared with the traditional technology, the microfluidic technology has a plurality of advantages in the aspect of analyzing samples: firstly, the micro-fluidic chip has flexible structural and functional design, and can meet the requirement of single-cell analysis. Second, typical microfluidic channels have dimensions of tens to hundreds of microns, can handle solution volumes ranging from picoliters to nanoliters, thereby reducing sample loss and high sensitivity, and enabling high throughput single cell analysis. In addition, the integration of the multifunctional unit and the microfluidic chip can realize automation and prevent measurement errors caused by manual operation.
Most microfluidic devices use microfluidic principles to isolate individual cells, hydrodynamic cell traps, pneumatic membrane valves, and oil drop based isolation. The most popular microfluidic isolation method at present is to use microdroplets to encapsulate single cells in inert carrier oil to form a closed space, thereby reducing the risk of sample contamination. Water-in-oil droplets produced by using water as the dispersed phase and the oil phase as the continuous phase. This type of droplet tends to be hydrophobic channel material. PDMS is the most common chip processing material at present, is a low-temperature thermal polymerization curing polymer material, has very strong shaping after shaping, has certain tolerance to solvents, and has the advantages of low price, good optical transparency, good biocompatibility, air permeability, convenient operation, high efficiency and the like when being used for manufacturing chips. Therefore, a series of means for detecting samples of the microfluidic chip can be developed based on the characteristics, the bonding mode of the PDMS chip is various, the operation is simple and convenient, simple reversible bonding can be realized by utilizing physical action, the chip can be reused, and irreversible bonding can also be realized by utilizing methods such as chemical modification and the like. However, higher hydrophobicity is required to form stable water-in-oil droplets, unmodified PDMS microfluidic chips are insufficient to provide such high hydrophobicity, and PDMS microfluidic chips also suffer from channel collapse, low loadings, and the like. Surface modification and finishing are required for application. .
The modification method of the polymer material can be generally adopted, and the modification method is divided into the following steps: surface modification and bulk modification, wherein the surface modification is most widely used and is divided into two major types, namely a physical modification and a chemical modification.
The chemical modification method can be classified into: wet modification and surface grafting by covalent bonding. The wet modification is to directly contact the solution to be modified with the surface of PDMS, so that the components to be modified on the surface of PDMS are adsorbed on the surface of PDMS through physical adsorption or electrostatic force. Common wet modification includes layer-by-layer self-assembly, sol-gel coating, dynamic surfactant modification, protein adsorption and the like. The common characteristic of the modification methods is that the modification methods are generally simpler. However, the PDMS surface and the modification layer are not connected by covalent chemical bonds, so that the modification layer has poor stability and is easy to run off along with the increase of the service time. The surface modification by covalent bonding is to covalently bond the modification layer to the PDMS surface by chemical reaction. If the modifying layer is also a type of polymer, such surface modification is also referred to as surface grafting. The greatest advantage of the modification method is that the modification layer is stable, the surface property after modification has long retention time, and the method is a more common method for chemical modification of PDMS chips.
The surface of PDMS is modified by Polydopamine (PDA), and the polydopamine is polymerized on the inner surface of a channel of the PDMS microfluidic chip under alkaline conditions to form a layer of PDA coating, so that the surface of the PDMS microfluidic chip is modified. However, the method has certain problems such as difficult polymerization operation in the microchannel, uneven coating thickness, unstable performance of the modified microfluidic chip, complicated preparation steps and the like.
In addition, MOFs are porous crystal materials formed by self-assembling metal ions or metal clusters and organic ligands through coordination bonds, can be used for depositing on the surface of a solid matrix or synthesizing proper micro-level coarse structures formed by MOFs particles, enriches the roughness of the surface of the materials, and can modify low-surface-energy chemical substances to enhance the surface hydrophobicity. The existing PDMS microfluidic chip channel surface modification mode is not easy to form a stable modification layer, so that MOFs are not applied to the surface modification of the PDMS microfluidic chip.
Disclosure of Invention
The invention provides a preparation method of a PDMS-PDA-MOFs micro-fluidic chip for single cell sequencing, which solves the problem of application of PDMS in the field of micro-fluidic chips.
The invention adopts the following technical scheme: a preparation method of PDMS-PDA-MOFs micro-fluidic chip for single cell sequencing comprises the steps of preparing an amino modified PDMS substrate; under alkaline condition, self-polymerizing DA (dopamine) solution on an amino modified PDMS substrate to form a layer of PDA film with super-strong adhesiveness, obtaining a PDMS substrate modified by PDA (polydopamine), and self-assembling and growing MOFs on the surface of the PDMS substrate to obtain a microsphere type PDMS-PDA-MOFs elastomer with an internal and external cladding microstructure; and curing, attaching and baking the PDMS-PDA-MOFs elastomer in a mold to finally obtain the PDMS-PDA-MOFs microfluidic chip.
Further, the method comprises the following steps:
(1) Preparation of amino-modified PDMS substrates: mixing vinyl silicone oil and PDMS according to a mass ratio of 2:1, adding 0.5g, heating at 50 ℃ for 1h, and activating to obtain a PDMS substrate containing alkenyl; dissolving 0.1g of cysteamine and 0.005g of dimethylolpropionic acid in 20mL of ethanol to form a solution A, dissolving a PDMS substrate containing alkenyl in the solution A, and reacting for 0.5-2h under ultraviolet light; washing with ethanol to remove unreacted substances, thereby obtaining an amino modified PDMS substrate;
(2) PDA modification of amino modified PDMS substrates: preparing a DA (dopamine) solution with the concentration of 2mg/ml by using a Tris hydrochloric acid solution with the pH of 8.5, pouring the DA (dopamine) solution into the amino modified PDMS substrate obtained in the step 1, uniformly mixing the DA (dopamine) solution, pouring the amino modified PDMS substrate into the space of 1-2ml, then placing the mixture in an environment of 25 ℃ for 24 hours, uniformly coating the PDA on the surface of the amino modified PDMS substrate by virtue of covalent bond bonding, washing the PDMS substrate by deionized water for several times due to the fact that the PDMS has lower surface energy, and washing away DA (dopamine) which is not adhered and is not firmly adhered on the PDMS to obtain the PDMS substrate modified by the PDA (polydopamine);
(3) Preparation of PDMS-PDA-MOFs elastomer: adding MOFs powder into a solvent for ultrasonic dispersion to obtain MOFs suspension, adding the MOFs suspension into a PDMS substrate modified by PDA (polydopamine) to promote the MOFs to self-assemble and grow on the surface of the PDMS substrate modified by the PDA (polydopamine), and placing the PDMS substrate in a 37 ℃ environment for 20 hours to obtain the PDMS-PDA-MOFs elastomer;
(4) PDMS-PDA-MOFs with micro channels were prepared: placing a silicon wafer with a mould in a clean glass dish, mixing a PDMS-PDA-MOFs elastic agent and a curing agent according to the mass ratio of 10:1 to prepare a mixed solution, pouring the mixed solution into the glass dish, removing bubbles in vacuum in a vacuum drying box for 1h at 90 ℃ for 2h, and cooling the PDMS-PDA-MOFs for curing; after stripping, PDMS-PDA-MOFs with micro-channels are obtained;
(5) Preparation of PDMS-PDA-MOFs microfluidic chips: placing a silicon wafer in a clean glass dish, mixing a PDMS-PDA-MOFs elastic agent and a curing agent according to the mass ratio of 10:1 to prepare a mixed solution, pouring the mixed solution into the glass dish, removing bubbles in vacuum in a vacuum drying box for 1h, baking the dried product at 90 ℃ for 2h, cooling the PDMS-PDA-MOFs, curing the cooled product to prepare a flat PDMS-PDA-MOFs, tightly attaching the flat PDMS-PDA-MOFs and the PDMS-PDA-MOFs with micro channels together, placing the flat PDMS-PDA-MOFs and the PDMS-MOFs into an oven, baking the flat PDMS-PDA-MOFs in the oven for 1h at 90 ℃, and taking out the flat PDMS-PDA-MOFs micro-fluidic chip after cooling.
Further, in the step (1), the ultraviolet light has a wavelength of 365nm and an effective distance of 10cm.
Further, the solid-to-liquid ratio of the MOFs powder and the solvent in step (3) is 1g: 80-120 ml.
Further, the curing agent is DBP (dibutyl phthalate).
Further, the solvent in the step (3) is dichloromethane (dichloromethane, CH2Cl 2), ethanol (ethanol, C2H5 OH) or dimethylformamide (n, n-dimethylformamide, DMF).
Further, the ultrasonic dispersion time in the step (3) is 45-75 min.
Further, the volume ratio of the MOFs suspension to the PDA (polydopamine) -modified PDMS substrate in step (3) is 1: 15-25.
Further, the cooling time in the step (4) is 1.5-2.5 h.
Further, the MOFs are ZIF-8, uiO-66 or UiO-67.
Compared with the prior art, the preparation method of the PDMS-PDA-MOFs micro-fluidic chip for single cell sequencing has the beneficial effects that:
(1) PDMS-PDA-MOFs elastomer is prepared by using a PDA template guiding method, and the PDA can guide the assembly of PDMS and MOFs to coat the MOFs. And the PDA is used for modifying the PDMS substrate material, and DA can be self-polymerized on the surface of PDMS particles in a solvent environment to form a layer of PDA film with super-strong adhesion. In addition, the ortho-position of the phenolic hydroxyl of the PDA can react with hydroxyl or sulfhydryl, amino and metal ions can be captured and chelated to form a nuclear growth site, so that strong adhesive force and secondary reaction capacity are provided for the subsequent modification of the PDMS surface, the modified PDMS surface can be used as a secondary platform, the self-assembly growth of MOFs ligand on the particle surface is promoted, the self-nucleation of MOFs is avoided, and the coating of the MOFs on the PDMS surface is realized.
(2) And the micro-fluidic chip is prepared by pouring the PDMS with the surface modified by adopting an integral method, and the preparation method is simple.
(3) The method comprises the steps of inducing a click chemistry method of sulfhydryl and olefin to react with cysteamine through ultraviolet irradiation to obtain the PDMS surface containing amino, and grafting PDA onto an amino modified PDMS substrate through Schiff base reaction/Michael addition reaction under the alkaline condition of room temperature so as to ensure uniform coating of the PDA.
(4) The coated microstructure ensures the stability of performance, and the coated microsphere structure forms a homogeneous PDMS-PDA-MOFs elastic agent, so that the structural instability caused by uneven coating due to macroscopic surface treatment can be avoided.
Drawings
FIG. 1 is a schematic diagram of the microstructure of a PDMS-PDA-MOFs material of the present invention used in the method of preparing a PDMS-PDA-MOFs microfluidic chip for single cell sequencing;
Fig. 2 is a schematic diagram of a chip channel of a microfluidic chip according to a method for preparing PDMS-PDA-MOFs microfluidic chips for single-cell sequencing according to the present invention.
Detailed Description
In order to further describe the technical means and effects adopted by the present invention for achieving the intended purpose, the following detailed description will refer to the specific implementation, structure, characteristics and effects according to the present invention with reference to the accompanying drawings and preferred embodiments.
Example 1
A preparation method of PDMS-PDA-MOFs micro-fluidic chip for single cell sequencing comprises the steps of firstly introducing DA solution into a channel of reaction equipment, and forming a PDA film to be adhered to the surface of PDMS under the self-polymerization of the DA solution in the channel under the alkaline condition; MOFs were further modified.
The method comprises the following steps:
(1) Preparation of amino-modified PDMS substrates: mixing vinyl silicone oil and PDMS (mass ratio 2:1), heating at 50deg.C for 1 hr, and activating to obtain PDMS substrate containing alkenyl; cysteamine (0.1 g) and dimethylolpropionic acid (0.005 g) were dissolved in ethanol (20 mL) to form solution A, and then PDMS substrate containing alkenyl group was dissolved in solution A for reaction under UV light for 0.5-2h; washing with ethanol to remove unreacted substances, thereby obtaining an amino modified PDMS substrate;
(2) PDA modification of amino modified PDMS substrates: preparing a DA (dopamine) solution with the concentration of 2mg/ml by using a Tris hydrochloric acid solution with the pH of 8.5, pouring the DA (dopamine) solution into the amino modified PDMS substrate obtained in the step 1, uniformly mixing the DA (dopamine) solution, pouring the amino modified PDMS substrate into the volume of 1ml, then placing the mixture in an environment of 25 ℃ for 24 hours, uniformly coating the PDA on the surface of the amino modified PDMS substrate by virtue of covalent bond bonding, washing the PDMS with deionized water for several times due to the fact that the PDMS has lower surface energy, and washing away DA (dopamine) which is not adhered and is not firmly adhered on the PDMS to obtain the PDMS substrate modified by the PDA (polydopamine);
(3) Preparation of PDMS-PDA-MOFs elastomer: adding MOFs powder into a solvent for ultrasonic dispersion to obtain MOFs suspension, adding the MOFs suspension into a PDMS substrate modified by PDA (polydopamine), promoting the MOFs to self-assemble and grow on the surface of the PDMS substrate modified by the PDA (polydopamine), placing the PDMS substrate in a 37 ℃ environment, and reacting for 20 hours to obtain the PDMS-PDA-MOFs elastomer;
(4) PDMS-PDA-MOFs with micro channels were prepared: placing a silicon wafer with a mould in a clean glass dish, mixing a PDMS-PDA-MOFs (polydimethylsiloxane-metal oxide semiconductor field effect transistor) elastic agent and a curing agent according to the mass ratio of 10:1 to prepare a mixed solution, pouring the mixed solution into the glass dish, removing bubbles in vacuum in a vacuum drying oven for 1h at 90 ℃ for 2h, and cooling the PDMS-PDA-MOFs for curing; after stripping, PDMS-PDA-MOFs with micro-channels are obtained;
(5) Preparation of PDMS-PDA-MOFs microfluidic chips: placing a silicon wafer in a clean glass dish, mixing a PDMS-PDA-MOFs elastic agent and a curing agent according to the mass ratio of 10:1 to prepare a mixed solution, pouring the mixed solution into the glass dish, pouring the mixed solution into a volume of 0.5ml, removing bubbles in vacuum in a vacuum drying box for 1h, baking the dried product at 90 ℃ for 2h, cooling the PDMS-PDA-MOFs to cure the dried product, preparing a flat PDMS-PDA-MOFs, tightly attaching the flat PDMS-PDA-MOFs and the PDMS-PDA-MOFs with micro-channels together, placing the flat PDMS-PDA-MOFs and the PDMS-MOFs into an oven, baking the flat PDMS-PDA-MOFs in the oven for 1h at 90 ℃, cooling the flat PDMS-PDA-MOFs micro-fluidic chip is obtained.
Finally, a latex tube is inserted into the inlet of a micro-channel of the PDMS-PDA-MOFs micro-fluidic chip, the inner diameter of the latex tube is 0.38mm, and the outer diameter of the latex tube is 1.09mm.
In this embodiment, the ultraviolet light in step (1) has a wavelength of 365nm and an effective distance of 10cm.
In this example, the solid-to-liquid ratio of MOFs powder to solvent in step (3) was 1g:80ml.
In this example, the curing agent is DBP (dibutyl phthalate).
In this example, the solvent in step (3) is methylene chloride (dichloromethane, CH2Cl 2), and in some examples, ethanol (ethanol, C2H5 OH) or dimethylformamide (n, n-dimethylformamide, DMF) may be used.
In this embodiment, the ultrasonic dispersion time in the step (3) is 45min.
In this example, the volume ratio of MOFs suspension to PDA (polydopamine) -modified PDMS substrate in step (3) was 1:15.
In this embodiment, the cooling time in step (4) is 1.5.
In this embodiment, MOFs are ZIF-8, and in some embodiments UiO-66 or UiO-67 may be used.
The water and oil contact angles of the prepared PDMS-PDA-MOFs surface are characterized by adopting a contact angle measuring instrument, so that the wettability of the PDMS-PDA-MOFs is analyzed. The specific process is as follows: a drop of 5 microliters of deionized water was dropped onto the PDMS-PDA-MOFs surface, immediately photographed using a water contact angle meter and the water contact angle values were calculated for each material, and 5 different points were tested for each material, as shown in table 1, with the calculated average value of the contact angle of the material, which was 141.7 °, indicating its superhydrophobicity.
Table 1 contact angle test
Example two
A preparation method of PDMS-PDA-MOFs micro-fluidic chip for single cell sequencing comprises the steps of firstly introducing DA solution into a channel of reaction equipment, and forming a PDA film to be adhered to the surface of PDMS under the self-polymerization of the DA solution in the channel under the alkaline condition; MOFs were further modified.
The method comprises the following steps:
(1) Preparation of amino-modified PDMS substrates: mixing vinyl silicone oil and PDMS (mass ratio 2:1), heating at 50deg.C for 1 hr, and activating to obtain PDMS substrate containing alkenyl; cysteamine (0.1 g) and dimethylolpropionic acid (0.005 g) were dissolved in ethanol (20 mL) to form a solution a, and then a PDMS substrate containing alkenyl groups was dissolved in the solution a and reacted under uv light for 2 hours; washing with ethanol to remove unreacted substances, thereby obtaining an amino modified PDMS substrate;
(2) PDA modification of amino modified PDMS substrates: preparing a DA (dopamine) solution with the concentration of 2mg/ml by using a Tris hydrochloric acid solution with the pH of 8.5, pouring the DA (dopamine) solution into the amino modified PDMS substrate obtained in the step 1, uniformly mixing the DA (dopamine) solution, pouring the amino modified PDMS substrate into the volume of 2ml, then placing the PDMS substrate in a 25 ℃ environment, reacting for 24 hours, uniformly coating the PDA on the surface of the amino modified PDMS substrate by virtue of covalent bond bonding, washing the PDMS substrate by deionized water for several times due to the fact that the PDMS has lower surface energy, and washing away DA (dopamine) which is not adhered and is not firmly adhered on the PDMS to obtain the PDMS substrate modified by the PDA (polydopamine);
(3) Preparation of PDMS-PDA-MOFs elastomer: adding MOFs powder into a solvent for ultrasonic dispersion to obtain MOFs suspension, adding the MOFs suspension into a PDMS substrate modified by PDA (polydopamine), promoting the MOFs to self-assemble and grow on the surface of the PDMS substrate modified by the PDA (polydopamine), placing the PDMS substrate in a 37 ℃ environment, and reacting for 20 hours to obtain the PDMS-PDA-MOFs elastomer;
(4) PDMS-PDA-MOFs with micro channels were prepared: placing a silicon wafer with a mould in a clean glass dish, mixing a PDMS-PDA-MOFs (polydimethylsiloxane-metal oxide semiconductor field effect transistor) elastic agent and a curing agent according to the mass ratio of 10:1 to prepare a mixed solution, pouring the mixed solution into the glass dish, removing bubbles in vacuum in a vacuum drying oven for 1h, baking the silicon wafer at 90 ℃ for 2h, and cooling the PDMS-PDA-MOFs for curing; after stripping, PDMS-PDA-MOFs with micro-channels are obtained;
(5) Preparation of PDMS-PDA-MOFs microfluidic chips: placing a silicon wafer in a clean glass dish, mixing a PDMS-PDA-MOFs elastic agent and a curing agent according to the mass ratio of 10:1 to prepare a mixed solution, pouring the mixed solution into the glass dish, pouring the mixed solution into a volume of 1ml, removing bubbles in vacuum in a vacuum drying oven for 1h, baking at 90 ℃ for 2h, cooling the PDMS-PDA-MOFs to cure, preparing flat PDMS-PDA-MOFs, tightly attaching the flat PDMS-PDA-MOFs and the PDMS-PDA-MOFs with micro channels together, placing the PDMS-PDA-MOFs and the curing agent in an oven, baking the PDMS-PDA-MOFs in the oven for 1h at 90 ℃, cooling, and taking out the PDMS-PDA-MOFs micro-fluidic chip.
Finally, a latex tube is inserted into the inlet of a micro-channel of the PDMS-PDA-MOFs micro-fluidic chip, the inner diameter of the latex tube is 0.38mm, and the outer diameter of the latex tube is 1.09mm.
In this embodiment, the ultraviolet light in step (1) has a wavelength of 365nm and an effective distance of 10cm.
In this example, the solid-to-liquid ratio of MOFs powder to solvent in step (3) was 1g:120ml.
In this example, the curing agent is DBP (dibutyl phthalate).
In this example, the solvent in step (3) is dimethylformamide (n, n-dimethylformamide, DMF), and in some examples, ethanol (ethanol, C2H5 OH) or dichloromethane (dichloromethane, CH2Cl 2) may be used.
In this embodiment, the ultrasonic dispersion time in step (3) is 75 minutes.
In this example, the volume ratio of MOFs suspension to PDA (polydopamine) -modified PDMS substrate in step (3) was 1:25.
In this example, the cooling time in step (4) was 2.5h.
In this embodiment, MOFs are UiO-67, and in some embodiments UiO-66 or ZIF-8 may be used.
The water and oil contact angles of the prepared PDMS-PDA-MOFs surface are characterized by adopting a contact angle measuring instrument, so that the wettability of the PDMS-PDA-MOFs is analyzed. The specific process is as follows: a drop of 5 microliters of deionized water was dropped onto the PDMS-PDA-MOFs surface, immediately photographed using a water contact angle meter and the water contact angle values were calculated for each material, and 5 different points were tested for each material, as shown in table 2, with the calculated average contact angle for the material, which was 152.7 °, indicating superhydrophobicity.
Table 2 contact angle test
The present invention is not limited to the above embodiments, but is capable of modification and variation in detail, and other modifications and variations can be made by those skilled in the art without departing from the scope of the present invention.
Claims (9)
1. A preparation method of a PDMS-PDA-MOFs micro-fluidic chip for single cell sequencing is characterized by firstly preparing an amino modified PDMS substrate; under alkaline condition, self-polymerizing DA (dopamine) solution on an amino modified PDMS substrate to form a layer of PDA film with super-strong adhesiveness, obtaining a PDMS substrate modified by PDA (polydopamine), and self-assembling and growing MOFs on the surface of the PDMS substrate to obtain a microsphere type PDMS-PDA-MOFs elastomer with an internal and external cladding microstructure; curing, attaching and baking the PDMS-PDA-MOFs elastomer in a mold to finally obtain the PDMS-PDA-MOFs micro-fluidic chip, which comprises the following steps:
(1) Preparation of amino-modified PDMS substrates: mixing vinyl silicone oil and PDMS (mass ratio 2:1), heating at 50deg.C for 1 hr, and activating to obtain PDMS substrate containing alkenyl; dissolving 0.1g of cysteamine and 0.005g of dimethylolpropionic acid in 20mL of ethanol to form a solution A, dissolving a PDMS substrate containing alkenyl in the solution A, and reacting for 0.5-2h under ultraviolet light; washing with ethanol to remove unreacted substances, thereby obtaining an amino modified PDMS substrate;
(2) PDA modification of amino modified PDMS substrates: preparing a DA (dopamine) solution with the concentration of 2mg/ml by using a Tris hydrochloric acid solution with the pH of 8.5, pouring the DA (dopamine) solution into the amino modified PDMS substrate obtained in the step 1, uniformly mixing the DA (dopamine) solution, pouring the amino modified PDMS substrate into the space of 1-2ml, then placing the mixture in an environment of 25 ℃ for 24 hours, uniformly coating the PDA on the surface of the amino modified PDMS substrate by virtue of covalent bond bonding, washing the PDMS substrate by deionized water for several times due to the fact that the PDMS has lower surface energy, and washing away DA (dopamine) which is not adhered and is not firmly adhered on the PDMS to obtain the PDMS substrate modified by the PDA (polydopamine);
(3) Preparation of PDMS-PDA-MOFs elastomer: adding MOFs powder into a solvent for ultrasonic dispersion to obtain MOFs suspension, adding the MOFs suspension into a PDMS substrate modified by PDA (polydopamine) to promote the MOFs to self-assemble and grow on the surface of the PDMS substrate modified by the PDA (polydopamine), and placing the PDMS substrate in a 37 ℃ environment for 20 hours to obtain the PDMS-PDA-MOFs elastomer;
(4) PDMS-PDA-MOFs with micro channels were prepared: placing a silicon wafer with a mould in a clean glass dish, mixing a PDMS-PDA-MOFs elastic agent and a curing agent according to the mass ratio of 10:1 to prepare a mixed solution, pouring the mixed solution into the glass dish, removing bubbles in vacuum in a vacuum drying box for 1h at 90 ℃ for 2h, and cooling the PDMS-PDA-MOFs for curing; after stripping, PDMS-PDA-MOFs with micro-channels are obtained;
(5) Preparation of PDMS-PDA-MOFs microfluidic chips: placing a silicon wafer in a clean glass dish, mixing a PDMS-PDA-MOFs elastic agent and a curing agent according to the mass ratio of 10:1 to prepare a mixed solution, pouring the mixed solution into the glass dish, removing bubbles in vacuum in a vacuum drying box for 1h, baking the dried product at 90 ℃ for 2h, cooling the PDMS-PDA-MOFs, curing the cooled product to prepare a flat PDMS-PDA-MOFs, tightly attaching the flat PDMS-PDA-MOFs and the PDMS-PDA-MOFs with micro channels together, placing the flat PDMS-PDA-MOFs and the PDMS-MOFs into an oven, baking the flat PDMS-PDA-MOFs in the oven for 1h at 90 ℃, and taking out the flat PDMS-PDA-MOFs micro-fluidic chip after cooling.
2. The method for preparing a PDMS-PDA-MOFs micro-fluidic chip for single cell sequencing according to claim 1 wherein the ultraviolet light in the step (1) is 365nm in wavelength and has an effective distance of 10cm.
3. The method for preparing a PDMS-PDA-MOFs microfluidic chip for single-cell sequencing according to claim 1, wherein the solid-to-liquid ratio of the MOFs powder and the solvent in the step (3) is 1g: 80-120 ml.
4. The method for preparing a PDMS-PDA-MOFs micro-fluidic chip for single cell sequencing according to claim 1 wherein the curing agent is DBP (dibutyl phthalate).
5. A method for preparing a PDMS-PDA-MOFs microfluidic chip for single cell sequencing according to claim 1 or 3, wherein the solvent in step (3) is dichloromethane (dichloromethane, CH2Cl 2), ethanol (ethanol, C2H5 OH) or dimethylformamide (n, n-dimethylformamide, DMF).
6. The method for preparing a PDMS-PDA-MOFs micro-fluidic chip for single cell sequencing according to claim 1, wherein the ultrasonic dispersion time in the step (3) is 45-75 min.
7. The method for preparing a PDMS-PDA-MOFs microfluidic chip for single cell sequencing according to claim 1, wherein the volume ratio of the MOFs suspension to the PDA (polydopamine) -modified PDMS substrate in the step (3) is 1: 15-25.
8. The method for preparing a PDMS-PDA-MOFs micro-fluidic chip for single cell sequencing according to claim 1, wherein the cooling time in the step (4) is 1.5-2.5 h.
9. The method for preparing a PDMS-PDA-MOFs micro-fluidic chip for single cell sequencing according to claim 1 wherein the MOFs are ZIF-8, uiO-66 or UiO-67.
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