CN115254209A - Preparation method of PDMS-PDA-MOF micro-fluidic chip for single cell sequencing - Google Patents
Preparation method of PDMS-PDA-MOF 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 by firstly preparing an amino modified PDMS substrate; carrying out self-polymerization on a DA (dopamine) solution on an amino modified PDMS substrate under an alkaline condition to obtain a PDA (polydopamine) modified PDMS substrate; modifying MOFs on the surface of the composite material to obtain a microspheric PDMS-PDA-MOFs elastic agent with a microstructure which is internally and externally coated; and curing, laminating and baking the PDMS-PDA-MOFs elastic agent 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 microfluidic chip, in particular to a preparation method of a PDMS-PDA-MOFs microfluidic chip for single cell sequencing.
Background
The heterogeneity of individual cells in a cell population plays a major role in the development and progression of disease, but most traditional genetic analysis methods currently mask individual cell differences. Single cell sequencing can exhibit intrinsic heterogeneity of individual cells and reveal complex and rare cell populations. In recent years, different microfluidic technologies have appeared for single cell research, becoming the leading edge of the field.
Compared to conventional techniques, microfluidic techniques have several advantages in analyzing samples: firstly, the structure and the function of the microfluidic chip are designed flexibly, and the requirement of single cell analysis can be met. Second, typical microfluidic channels have dimensions of tens to hundreds of microns, and can handle solution volumes 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 measurement errors caused by manual operation are prevented.
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 encapsulate the single cells in an inert carrier oil using microdroplets to form a closed space, reducing the risk of sample contamination. Water-in-oil droplets are generated by taking water as a dispersed phase and taking an oil phase as a continuous phase. This type of droplet tends to be a hydrophobic channel material. PDMS is the most common chip processing material at present, is a low-temperature thermal polymerization curing polymer material, has strong moulding after molding, 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 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 a physical effect, the chip can be repeatedly used, and irreversible bonding can also be realized by utilizing methods such as chemical modification. However, higher hydrophobicity is required to form stable water-in-oil droplets, an unmodified PDMS microfluidic chip is not sufficient to provide such high hydrophobicity, and the PDMS microfluidic chip also has the disadvantages of easy deformation and collapse of channels, low loading capacity, and the like. Surface modification and modification are required for application. .
Generally, a modification method of a high polymer material can be adopted, and the modification method is divided into the following modification ranges: surface modification methods and bulk modification methods, wherein the surface modification methods are most widely used and are subdivided into two main types, namely physical modification methods and chemical modification methods.
Chemical modification methods can be divided into: wet modification and surface grafting by covalent bonding. The wet modification is to make the solution to be modified contact the surface of PDMS directly, so that the component to be modified on the surface of PDMS is adsorbed on the surface of PDMS by physical adsorption or electrostatic force. Common wet modification methods include 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 simpler overall. However, since the PDMS surface is not linked to the modification layer by covalent chemical bonds, the modification layer has poor stability and is easily lost with the increase of the usage time. The surface modification by covalent bond is to bond the modification layer to the PDMS surface by covalent bond through chemical reaction. If the modification layer is also a type of polymer, such surface modification is also referred to as surface grafting. The modification method has the greatest advantages that the modification layer is stable, the surface property after modification is long in retention time, and the method is a common method for chemical modification of the PDMS chip.
In the prior art, poly-dopamine (PDA) is used to modify the surface of PDMS, and dopamine is self-polymerized into poly-dopamine on the inner surface of the PDMS microfluidic chip channel under an alkaline condition to form a layer of PDA coating, thereby modifying the surface of the PDMS microfluidic chip. 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, the 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 MOFs particles later to form a proper micro-level rough structure, enriches the roughness of the surface of the material, and can modify low-surface-energy chemical substances to enhance the surface hydrophobic property. The existing PDMS microfluidic chip channel surface modification method is not easy to form a stable modification layer, so that MOFs are not applied to surface modification of PDMS microfluidic chips.
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 preparing amino modified PDMS substrate; carrying out self-polymerization on DA (dopamine) solution on an amino modified PDMS substrate under an alkaline condition to form a layer of PDA film with super strong adhesion, obtaining a PDMS substrate modified by PDA (poly-dopamine), and then growing MOFs on the surface of the PDMS substrate in a self-assembly manner to obtain a microspheric PDMS-PDA-MOFs elastic agent with a microstructure coated inside and outside; and curing, laminating and baking the PDMS-PDA-MOFs elastic agent 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 substrate: mixing vinyl silicone oil and PDMS according to a mass ratio of 2 (1), heating for 1h at 50 ℃ for activation by 0.5g in total to obtain a PDMS substrate containing alkenyl; dissolving cysteamine (0.1 g) and dimethylolpropionic acid (0.005 g) in ethanol (20 mL) to form a solution A, then 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 to obtain an amino modified PDMS substrate;
(2) Performing PDA modification on the amino modified PDMS substrate: preparing a DA (dopamine) solution with the concentration of 2mg/ml by using a Tris hydrochloric acid solution with the pH =8.5, pouring the solution into the amino modified PDMS substrate obtained in the step 1, uniformly mixing, pouring the mixture into a volume of 1-2ml, then placing the mixture in an environment at 25 ℃, acting for 24 hours, and combining the mixture by virtue of a covalent bond, wherein the PDMS has lower surface energy, so that the PDA is uniformly coated on the surface of the amino modified PDMS substrate, washing the PDMS substrate for several times by using deionized water, and washing off 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) Preparing a PDMS-PDA-MOFs elastic agent: adding MOFs powder into a solvent for ultrasonic dispersion to obtain an MOFs suspension, adding the MOFs suspension into a PDA (poly dopamine) -modified PDMS substrate to promote the self-assembly growth of the MOFs on the surface of the PDA (poly dopamine) -modified PDMS substrate, placing the substrate in an environment at 37 ℃, and acting for 20 hours to obtain a PDMS-PDA-MOFs elastic agent;
(4) Preparing PDMS-PDA-MOFs with micro-channels: placing a silicon wafer with a mould in a clean glass dish, mixing a PDMS-PDA-MOFs (polydimethylsiloxane-PDA-metal organic frameworks) elastic agent and a curing agent in a mass ratio of 10; obtaining PDMS-PDA-MOFs with the micro-channel after stripping;
(5) Preparing a PDMS-PDA-MOFs microfluidic chip: placing a silicon wafer in a clean glass dish, mixing a PDMS-PDA-MOFs elastic agent and a curing agent according to a mass ratio of 10.
Further, the wavelength of the ultraviolet light in the step (1) is 365nm, and the effective distance is 10cm.
Further, the solid-to-liquid ratio of the MOFs powder to the solvent in the step (3) is 1g: (80-120) ml.
Further, the curing agent is DBP (dibutyl phthalate).
Further, in the step (3), the solvent is dichloromethane (CH 2Cl 2), ethanol (ethanol, C2H5 OH) or Dimethylformamide (DMF).
Further, the ultrasonic dispersion time in the step (3) is 45-75 min.
Further, the volume ratio of the MOFs suspension liquid to the PDA (poly dopamine) -modified PDMS substrate in the step (3) is 1: (15 to 25).
Further, the cooling time in the step (4) is 1.5-2.5 h.
Furthermore, 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 following beneficial effects:
(1) And preparing the PDMS-PDA-MOFs elastic agent by utilizing a PDA template guiding method, wherein the PDA can guide the assembly of PDMS and MOFs and coat the MOFs. The PDA is used for modifying the PDMS substrate material, and the 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 group of the PDA can react with the hydroxyl group or the sulfydryl group, the amino group, the capturable metal ion and the chelated metal ion can be used as a nuclear growth site, strong adhesive force and secondary reaction capacity are provided for subsequent modification of the surface of the PDMS, the PDMS can be used as a secondary platform, the MOFs ligand can be promoted to grow in a self-assembly mode on the particle surface, the MOFs are prevented from nucleating automatically, and the MOFs on the surface of the PDMS can be coated.
(2) The integrated method is adopted to cast the PDMS with the modified surface to prepare the micro-fluidic chip, and the preparation method is simple and convenient.
(3) The method comprises the steps of reacting ultraviolet light irradiation induced sulfydryl and olefin with cysteamine to obtain an amino-containing PDMS surface, and grafting PDA to an amino-modified PDMS substrate successfully through Schiff base reaction/Michael addition reaction under the room temperature alkaline condition to ensure the uniform coating of the PDA.
(4) The coating microstructure ensures the stability of performance, and the homogeneous PDMS-PDA-MOFs elastic agent exists in the coating microsphere structure, so that the structural instability caused by uneven coating due to macroscopic surface treatment can be avoided.
Drawings
FIG. 1 is a schematic view of the microstructure of PDMS-PDA-MOFs material of the preparation method of PDMS-PDA-MOFs microfluidic chip for single cell sequencing according to the present invention;
FIG. 2 is a schematic diagram of a chip channel of a microfluidic chip of the preparation method of the PDMS-PDA-MOFs microfluidic chip for single cell sequencing according to the present invention.
Detailed Description
To further illustrate the technical means and effects of the present invention adopted to achieve the predetermined objects, the following detailed description of the embodiments, structures, features and effects according to the present invention will be made with reference to the accompanying drawings and preferred embodiments.
Example one
A method for preparing PDMS-PDA-MOFs micro-fluidic chip for single cell sequencing comprises introducing DA solution into a channel, and allowing the DA solution to undergo self-polymerization in the channel under alkaline condition to form PDA film adhered to the surface of the PDMS channel; and modifying the upper MOFs.
The method comprises the following steps:
(1) Preparation of amino-modified PDMS substrate: mixing vinyl silicone oil and PDMS according to a mass ratio of 2 (1), heating for 1h at 50 ℃ for activation by 0.5g in total to obtain a PDMS substrate containing alkenyl; dissolving cysteamine (0.1 g) and dimethylolpropionic acid (0.005 g) in ethanol (20 mL) to form a solution A, then 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 to obtain an amino modified PDMS substrate;
(2) Modification of PDA on amino modified PDMS substrate: preparing a DA (dopamine) solution with the concentration of 2mg/ml by using a Tris hydrochloric acid solution with the pH =8.5, pouring the solution into the amino modified PDMS substrate obtained in the step 1, uniformly mixing, pouring the mixture into a volume of 1ml, then placing the mixture in an environment at 25 ℃, acting for 24 hours, and combining the mixture by virtue of a covalent bond, wherein the PDMS has lower surface energy, so that the PDA is uniformly coated on the surface of the amino modified PDMS substrate, washing the PDMS substrate for several times by using deionized water, and washing off the 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) Preparing a PDMS-PDA-MOFs elastic agent: adding MOFs powder into a solvent for ultrasonic dispersion to obtain an MOFs suspension, adding the MOFs suspension into a PDA (poly dopamine) -modified PDMS substrate to promote the self-assembly growth of the MOFs on the surface of the PDA (poly dopamine) -modified PDMS substrate, placing the substrate in an environment at 37 ℃, and acting for 20 hours to obtain a PDMS-PDA-MOFs elastic agent;
(4) Preparing PDMS-PDA-MOFs with microchannels: placing a silicon wafer with a mold in a clean glass dish, mixing a PDMS-PDA-MOFs (polydimethylsiloxane-metal-organic frameworks) elastic agent and a curing agent according to a mass ratio of 10; stripping to obtain PDMS-PDA-MOFs with microchannels;
(5) Preparing a PDMS-PDA-MOFs microfluidic chip: placing a silicon wafer in a clean glass dish, mixing a PDMS-PDA-MOFs (polydimethylsiloxane-PDA-metal-organic frameworks) elastic agent and a curing agent according to a mass ratio of 10 to prepare a mixed solution, then pouring the mixed solution into the glass dish, wherein the volume of the glass dish is 0.5ml, removing bubbles in a vacuum drying oven for 1h, baking the mixed solution at 90 ℃ for 2h, cooling the PDMS-PDA-MOFs for curing to obtain flat PDMS-PDA-MOFs, closely attaching the flat PDMS-PDA-MOFs to PDMS-PDA-MOFs with a microchannel, placing the PDMS-PDA-MOFs in an oven, baking the flat PDMS-PDA-MOFs at 90 ℃ for 1h, cooling and taking out to obtain the PDMS-PDA-MOFs microfluidic chip.
And finally, inserting a latex tube at the inlet of the micro-channel of the PDMS-PDA-MOFs microfluidic chip, wherein 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 wavelength of the ultraviolet light in step (1) is 365nm, and the effective distance is 10cm.
In this embodiment, the solid-to-liquid ratio of the MOFs powder to the solvent in step (3) is 1g:80ml.
In this example, the curing agent was DBP (dibutyl phthalate).
In this embodiment, the solvent in step (3) is dichloromethane (CH 2Cl 2), and in some embodiments, ethanol (ethanol, C2H5 OH) or dimethylformamide (n, n-dimethylformamide, DMF) may also be used.
In this example, the ultrasonic dispersion time in step (3) was 45min.
In this embodiment, the volume ratio of the MOFs suspension to the PDA (polydopamine) -modified PDMS substrate in step (3) is 1:15.
in this example, the cooling time in step (4) was 1.5.
In this embodiment, the MOFs are ZIF-8, and in some embodiments, uiO-66 or UiO-67 may also be used.
And (3) adopting a contact angle measuring instrument to represent the contact angles of water and oil on the surface of the prepared PDMS-PDA-MOFs so as to analyze the wettability of the PDMS-PDA-MOFs. The specific process is as follows: a drop of 5 microliters of deionized water was dropped on the PDMS-PDA-MOFs surface, a photograph was immediately taken with a water contact angle measuring instrument and the water contact angle value was calculated, and each material was tested for 5 different points, as shown in Table 1, and the calculated average value was the contact angle of the material, which was 141.7 degrees, indicating its superhydrophobicity.
Table 1 contact Angle test
Example two
A method for preparing PDMS-PDA-MOFs micro-fluidic chip for single cell sequencing comprises introducing DA solution into a channel, and allowing the DA solution to undergo self-polymerization in the channel under alkaline condition to form PDA film adhered to the surface of the PDMS channel; the upper MOFs were further modified.
The method comprises the following steps:
(1) Preparation of amino-modified PDMS substrate: mixing vinyl silicone oil and PDMS according to a mass ratio of 2 (1), heating for 1h at 50 ℃ for activation by 0.5g in total to obtain a PDMS substrate containing alkenyl; dissolving cysteamine (0.1 g) and dimethylolpropionic acid (0.005 g) in ethanol (20 mL) to form a solution A, then dissolving a PDMS substrate containing alkenyl in the solution A, and reacting for 2h under ultraviolet light; washing with ethanol to remove unreacted substances to obtain an amino modified PDMS substrate;
(2) Modification of PDA on amino modified PDMS substrate: preparing a DA (dopamine) solution with the concentration of 2mg/ml by using a Tris hydrochloric acid solution with the pH =8.5, pouring the solution into the amino modified PDMS substrate obtained in the step 1, uniformly mixing, pouring the solution into a volume of 2ml, then placing the solution in an environment at 25 ℃, acting for 24 hours, and combining the solution by virtue of a covalent bond, wherein the PDMS has lower surface energy, so that the PDA is uniformly coated on the surface of the amino modified PDMS substrate, washing the PDMS substrate for several times by using deionized water, and washing off the 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) Preparing a PDMS-PDA-MOFs elastic agent: adding MOFs powder into a solvent for ultrasonic dispersion to obtain an MOFs suspension, adding the MOFs suspension into a PDA (poly dopamine) -modified PDMS substrate to promote the self-assembly growth of the MOFs on the surface of the PDA (poly dopamine) -modified PDMS substrate, placing the substrate in an environment at 37 ℃, and acting for 20 hours to obtain a PDMS-PDA-MOFs elastic agent;
(4) Preparing PDMS-PDA-MOFs with micro-channels: placing a silicon wafer with a mold in a clean glass dish, mixing a PDMS-PDA-MOFs (polydimethylsiloxane-metal-organic frameworks) elastic agent and a curing agent according to a mass ratio of 10; stripping to obtain PDMS-PDA-MOFs with microchannels;
(5) Preparing a PDMS-PDA-MOFs microfluidic chip: 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.
And finally, inserting a latex tube at the inlet of the micro-channel of the PDMS-PDA-MOFs microfluidic chip, wherein 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 wavelength of the ultraviolet light in step (1) is 365nm, and the effective distance is 10cm.
In this example, the solid-to-liquid ratio of the MOFs powder to the solvent in step (3) is 1g:120ml of the solution.
In this example, the curing agent was DBP (dibutyl phthalate).
In this embodiment, the solvent in step (3) is Dimethylformamide (DMF), and in some embodiments, ethanol (ethanol, C2H5 OH) or dichloromethane (CH 2Cl 2) may also be used.
In this example, the ultrasonic dispersion time in step (3) was 75min.
In this embodiment, the volume ratio of the MOFs suspension to the PDA (polydopamine) -modified PDMS substrate in step (3) is 1:25.
in this example, the cooling time in step (4) was 2.5 hours.
In this embodiment, the MOFs is UiO-67, and in some embodiments, uiO-66 or ZIF-8 may also be used.
And (3) characterizing the contact angles of water and oil on the surface of the prepared PDMS-PDA-MOFs by using a contact angle measuring instrument so as to analyze the wettability of the PDMS-PDA-MOFs. The specific process is as follows: a drop of 5 microliters of deionized water was dropped on the PDMS-PDA-MOFs surface, a photograph was immediately taken with a water contact angle meter and the water contact angle value was calculated, 5 different points were measured for each material, and as shown in Table 2, the calculated average value was the contact angle of the material, which was 152.7 degrees, indicating its superhydrophobicity.
Table 2 contact Angle test
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (10)
1. A preparation method of PDMS-PDA-MOFs micro-fluidic chip for single cell sequencing is characterized in that firstly, an amino modified PDMS substrate is prepared; carrying out self-polymerization on DA (dopamine) solution on an amino modified PDMS substrate under an alkaline condition to form a layer of PDA film with super strong adhesion, obtaining a PDMS substrate modified by PDA (poly-dopamine), and then growing MOFs on the surface of the PDMS substrate in a self-assembly manner to obtain a microspheric PDMS-PDA-MOFs elastic agent with a microstructure coated inside and outside; and curing, laminating and baking the PDMS-PDA-MOFs elastic agent in a mold to finally obtain the PDMS-PDA-MOFs microfluidic chip.
2. The method for preparing PDMS-PDA-MOFs microfluidic chip for single cell sequencing according to claim 1, comprising the following steps:
(1) Preparation of amino-modified PDMS substrate: mixing vinyl silicone oil and PDMS (polydimethylsiloxane) according to a mass ratio of (2); dissolving cysteamine (0.1 g) and dimethylolpropionic acid (0.005 g) in ethanol (20 mL) to form a solution A, then 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 to obtain an amino modified PDMS substrate;
(2) Performing PDA modification on the amino modified PDMS substrate: preparing a DA (dopamine) solution with the concentration of 2mg/ml by using a Tris hydrochloric acid solution with the pH =8.5, pouring the solution into the amino modified PDMS substrate obtained in the step 1, uniformly mixing, pouring the solution into a volume of 1-2ml, then placing the mixture in an environment at 25 ℃, acting for 24 hours, and combining the solution by virtue of a covalent bond, wherein due to the fact that PDMS has low surface energy, PDA is uniformly coated on the surface of the amino modified PDMS substrate, washing the surface of the amino modified PDMS substrate for several times by using deionized water, and washing off the DA (dopamine) which is not adhered and is not firmly adhered on the PDMS to obtain the PDMS substrate modified by PDA (poly dopamine);
(3) Preparing a PDMS-PDA-MOFs elastic agent: adding MOFs powder into a solvent for ultrasonic dispersion to obtain an MOFs suspension, adding the MOFs suspension into a PDA (poly dopamine) -modified PDMS substrate to promote the self-assembly growth of the MOFs on the surface of the PDA (poly dopamine) -modified PDMS substrate, placing the substrate in an environment at 37 ℃, and acting for 20 hours to obtain a PDMS-PDA-MOFs elastic agent;
(4) Preparing PDMS-PDA-MOFs with micro-channels: placing a silicon wafer with a mold in a clean glass dish, mixing a PDMS-PDA-MOFs (polydimethylsiloxane-moving oxygen-silicon-free functional elastomers) and a curing agent according to a mass ratio of 10; stripping to obtain PDMS-PDA-MOFs with microchannels;
(5) Preparing a PDMS-PDA-MOFs microfluidic chip: placing a silicon wafer in a clean glass dish, mixing a PDMS-PDA-MOFs elastic agent and a curing agent according to a mass ratio of 10.
3. The method for preparing PDMS-PDA-MOFs microfluidic chip for single cell sequencing according to claim 2, wherein the wavelength of the ultraviolet light in step (1) is 365nm, and the effective distance is 10cm.
4. The method for preparing PDMS-PDA-MOFs microfluidic chip for single cell sequencing according to claim 2, wherein the solid-to-liquid ratio of said MOFs powder and said solvent in step (3) is 1g: (80-120) ml.
5. The method for preparing PDMS-PDA-MOFs microfluidic chip for single cell sequencing according to claim 2, wherein the curing agent is DBP (dibutyl phthalate).
6. The method for preparing a PDMS-PDA-MOFs microfluidic chip for single cell sequencing according to claim 2 or 4, wherein the solvent in step (3) is dichloromethane (CH 2Cl 2), ethanol (ethanol, C2H5 OH) or dimethylformamide (n, n-dimethylformamide, DMF).
7. The method for preparing PDMS-PDA-MOFs microfluidic chip for single cell sequencing according to claim 2, wherein the ultrasonic dispersion time in step (3) is 45-75 min.
8. The method for preparing PDMS-PDA-MOFs microfluidic chip for single cell sequencing according to claim 2, wherein the volume ratio of said MOFs suspension to the PDA (polydopamine) -modified PDMS substrate in step (3) is 1: (15 to 25).
9. The method for preparing PDMS-PDA-MOFs microfluidic chip for single cell sequencing according to claim 2, wherein the cooling time in step (4) is 1.5-2.5 h.
10. The method for preparing PDMS-PDA-MOFs microfluidic chip for single cell sequencing according to claim 1 or 2, wherein the MOFs is ZIF-8, uiO-66 or UiO-67.
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