Surfactant as well as preparation method and related application thereof

Abstract

The invention discloses a surfactant as well as a preparation method and related application thereof, and relates to the field of analysis and detection.The surfactant is obtained through a polymerization reaction of a compound 1 and a compound 2, the synthesis process is simple and rapid, few byproducts are produced, the yield is high, liquid drops stabilized by the surfactant have good stability and biocompatibility, and the surfactant can be applied to the field of chemical engineering. The method is suitable for most liquid drop technologies, and has wide application potential in the fields of analytical chemistry, clinical medicine, cell biology, immunology, molecular biology and the like.

Description

Technical Field

[0001] The present invention relates to the field of molecular detection, and in particular, to a surfactant, a preparation method thereof, and related applications. Background Art

[0002] Droplet-based microfluidics refers to a technology in which, at the microscale, two immiscible liquid phases use the fluid shear force of the continuous phase (oil phase) to break the surface tension of the discrete phase, and cut the discrete phase (aqueous phase) into nanoliter or even picoliter droplets. Due to its outstanding advantages, such as low cost, high throughput, and high degree of automation, droplet-based microfluidics has become a very powerful tool in the field of biochemical analysis and is widely used in multiple aspects such as digital PCR, material synthesis, single-molecule enzyme sorting, microbial culture, drug delivery, single-molecule detection, and single-cell sequencing. Surfactants are an essential part of maintaining droplet stability. The presence of surfactants can greatly reduce the interfacial tension of droplets and reduce the aggregation or fragmentation between droplets.

[0003] Existing surfactants such as PEG-PFPE 2 (a triblock fluorinated surfactant, where PEG is polyethylene glycol and PFPE is perfluoropolyether) have some deficiencies. For example, their stability and biocompatibility need to be improved, and the preparation process is cumbersome, etc.

[0004] In view of this, the present invention is specifically proposed. Summary of the Invention

[0005] The purpose of the present invention is to provide a surfactant, a preparation method thereof, and related applications.

[0006] The present invention is implemented as follows:

[0007] In a first aspect, the present invention provides a polymer obtained by copolymerizing compound 1 and compound 2, and the structural formula of compound 1 is as shown in formula 1;

[0008] Formula 1:

[0009]

[0010] Wherein, R 1 is O or NH, and R 2 is selected from any one of an alkyl group with 1 to 5 carbon atoms, a hydroxyl group, and CH 2 =CH-C(=O)-, and the r is an integer from 2 to 500, and s is an integer from 0 to 6;

[0011] The structural formula of compound 2 is as shown in formula 2;

[0012] Formula 2:

[0013]

[0014] Wherein, R 3 is O or NH, R 4 is CF 2 、CHF or CF 2 CHF, p is an integer from 1 to 6, and q is an integer from 1 to 30.

[0015] In a second aspect, the present invention provides a method for preparing the aforementioned polymer, which includes the step of polymerizing compound 1 and compound 2.

[0016] In a third aspect, the present invention provides the use of the aforementioned polymer as a surfactant in the preparation of a surfactant-containing product.

[0017] In a fourth aspect, the present invention provides a surfactant-containing product, wherein the surfactant includes the aforementioned polymer.

[0018] In a fifth aspect, the present invention provides the use of the polymer described in the aforementioned embodiments as a surfactant or the aforementioned surfactant-containing product in droplet microfluidics technology.

[0019] The present invention has the following beneficial effects:

[0020] (1) By copolymerizing specific compound 1 and specific compound 2 to obtain a surfactant, it has the advantages of simple and rapid synthesis process, few by-products, high yield, etc. Moreover, the synthesis process does not require complex large-scale instruments and can be prepared in a conventional laboratory, significantly reducing the preparation cost of the surfactant.

[0021] (2) The droplets stabilized by the surfactant prepared in this application have good thermal stability and are stable at 37 °C, 60 °C and 95 °C; the surfactant prepared in this application has good stability for droplets in different aqueous phases; the surfactant prepared in this application has good biocompatibility and can complete the proliferation of single bacteria and cells in the droplets, providing a reliable platform for subsequent single-cell and single-molecule detection.

[0022] (3) The surfactant prepared in this application is applicable to most droplet technologies and has broad application potential in the fields of analytical chemistry, clinical medicine, cell biology, immunology, molecular biology, etc. Description of the Drawings

[0023] To more clearly illustrate the technical solutions of the embodiments of the present invention, the following will briefly introduce the drawings required for the embodiments. It should be understood that the following drawings only show some embodiments of the present invention, and thus should not be regarded as limiting the scope. For those of ordinary skill in the art, without creative efforts, other related drawings can also be obtained based on these drawings.

[0024] Figure 1 Synthesis schematic diagram of the surfactant for Embodiment 1 of the present invention;

[0025] Figure 2 In which, A is the infrared spectrum of polyethylene glycol diacrylate (PEGDA); B is the infrared spectrum of hexafluorobutyl methacrylate (MMHFB); C is the infrared spectrum of the PEGDA-MMHFB copolymer;

[0026] Figure 3 is 1 1H NMR-PEGDA nuclear magnetic diagram;

[0027] Figure 4 is 1 1H NMR-MMHFB nuclear magnetic diagram;

[0028] Figure 5 is 19 19F NMR-MMHFB nuclear magnetic diagram;

[0029] Figure 6 is 1 1H NMR-PEGDA-MMHFB copolymer nuclear magnetic diagram;

[0030] Figure 7 is 19 19F NMR-PEGDA-MMHFB copolymer nuclear magnetic diagram;

[0031] Figure 8 In which, A is the CMC of the PEGDA-MMHFB copolymer based on the DLS method; B is the dynamic surface tension of the PEGDA-MMHFB copolymer;

[0032] Figure 9 Experiment for adjusting the droplet size; among them, (A) when the oil-water flow rate ratio distribution is 1:1, 5:1, 10:1, the average droplet diameters are 95μm, 78μm, and 55μm respectively; (B) chips with intersections of different sizes 20, 50, 80μm can generate droplets with a diameter range of 25, 70, 95μm, scale bar = 100μm;

[0033] Figure 10For the droplet generation stability experiment; among them, (A) Droplet images prepared with water, PBS, LB medium, and YPD medium as the dispersed phase, placed at room temperature for 0 and 48 hours, scale bar = 100 μm; (B) Calculation of the droplet diameter distribution graphs at 0 hour and 48 hours.

[0034] Figure 11 For the droplet time stability and thermal stability experiment; among them, (A) Droplet stability images before and after being placed at room temperature for 7 days; (B) Droplet diameter distribution graphs before and after being placed at room temperature for 7 days; (C) Stability of the droplets before and after PCR reaction; (D) Droplet diameter distribution graphs before and after PCR reaction, scale bar = 100 μm.

[0035] Figure 12 For the biocompatibility experiment; among them, (A) Growth images and fluorescence intensity statistical graphs of individual Escherichia coli in droplets; (B) Schematic diagram and diameter distribution graph showing the gradual decrease in the diameter of droplets containing individual yeast cells due to the growth of yeast cells.

[0036] Figure 13 For the thermal cycling stability of different molecular brush fluorosurfactants; among them, (A) Comparison of 3 molecular brush fluorosurfactants before and after thermal cycling; (B) Statistical analysis graph of droplet diameter distribution, scale bar = 100 μm.

[0037] Figure 14 For the time stability of droplets generated by different fluorosurfactants; among them, (A) Droplet images of 3 types of droplets before and after being placed at room temperature for 7 days; (B) Droplet diameter distribution graphs before and after being placed at room temperature for 7 days, scale bar = 100 μm. Detailed implementation manners

[0038] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. For those not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. For reagents or instruments not specified by the manufacturer, they are all conventional products that can be obtained through commercial purchase.

[0039] Fluorosurfactants usually contain a hydrophilic group at one end and a hydrophobic group at the other end. By changing the functional group types of the hydrophilic group or the hydrophobic group, different functions are imparted to the surfactants. Compared with other surfactants, fluorosurfactants have multiple carbon-fluorine bonds, and carbon-fluorine bonds have extremely high polarity and chemical stability in nature, and are suitable for various reaction environments. In addition, fluorosurfactant molecules are small and can be dissolved in a variety of oil phases, and are suitable for a variety of narrow microfluidic channels without causing blockage. As a commonly used surfactant in the past, PEG-PFPE 2The lipophilic end of the triblock fluorinated surfactant is a long fluorocarbon chain perfluoropolyether (PFPE), and the hydrophilic end is polyethylene glycol (PEG). It takes into account both stability and certain biocompatibility and is currently the most widely used surfactant. Although it can stabilize monodisperse droplets for a long time, it still has the following disadvantages: (1) Stability needs to be improved: When the droplets are subjected to high temperature heat treatment, the stability of the droplets will be reduced, causing the droplets to fuse; (2) The preparation is cumbersome: PEG-PFPE 2 The manufacturing process of triblock fluorinated surfactants is usually carried out in a harsh environment of strong acid or strong base. The reaction requires multiple steps, which consumes manpower and reagents but has a low yield. Many by-products are easily produced during the reaction. If these by-products are difficult to remove, they may interfere with or even fail a series of downstream analyses such as amplification reactions or microbial culture.

[0040] In view of the defects of the prior art, the present invention provides a new fluorinated surfactant, which is obtained by copolymerizing a compound 1 having a specific structural formula with a compound 2 having a specific structural formula. Compared with the existing preparation process of the fluorinated surfactant, the preparation process of the fluorinated surfactant of the present application is simple to operate, less time-consuming, and has the advantages of less by-products, high yield, low cost, easy storage and transportation; the surfactant provided by the present invention overcomes the technical difficulties of traditional fluorinated surfactants, and the droplets prepared using the surfactant of the present application have the advantages of high stability and good biocompatibility, providing a better way for the application of droplet microfluidics technology.

[0041] In one aspect, the present invention provides a polymer obtained by copolymerizing a compound 1 and a compound 2, wherein the structural formula of the compound 1 is shown in Formula 1;

[0042] Formula 1:

[0043]

[0044] Among them, R 1 is O or NH, R 2 Selected from: alkyl with 1 to 5 carbon atoms, hydroxyl, CH 2 =CH-C(=O)-, wherein r is an integer of 2 to 500, and s is an integer of 0 to 6;

[0045] The structural formula of the compound 2 is shown in Formula 2;

[0046] Formula 2:

[0047]

[0048] Among them, R 3 is O or NH, R 4 For CF 2, CHF or CF 2 CHF, where p is an integer from 1 to 6 and q is an integer from 1 to 30.

[0049] In some embodiments, the polymerization is C═C polymerization.

[0050] Compound 1 has a hydrophilic head group and a hydrophilic tail group, and compound 2 has a hydrophobic head group and a hydrophobic tail group. Compound 1 and compound 2 can be copolymerized under the condition of a thermal initiator to obtain a polymer (molecular brush type), which can be referred to Figure 1 for the schematic diagram of Figure 1 Taking polyethylene glycol diacrylate PEGDA as compound 1 and 1H,1H,2H,2H-hexfluorobutyl methacrylate MMHFB as compound 2 as an example, after polymerization, a polymer is formed, and the structure is as follows:

[0051]

[0052] The final polymer may be one of these two polymer monomers or a mixture.

[0053] In some embodiments, the number of carbon atoms of the alkyl group with 1 to 5 carbon atoms is any one of 1, 2, 3, 4, and 5.

[0054] In some embodiments, r is any one of 2, 4, 6, 8, 10, 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, 490, 492, 494, 496, and 500 or the range between any two of them.

[0055] In some embodiments, the molecular weight of the polyethylene glycol group in formula 1 is 50 to 10,000. In some embodiments, the molecular weight of the polyethylene glycol group is any one of 50, 60, 70, 80, 90, 100, 500, 1000, 2000, 3000, 4000, 5000, 6000, 7000, 8000, 9000, 9900, 9910, 9920, 9930, 9940, 9950, 9960, 9970, 9980, 9990, and 10,000 or the range between any two of them, and is a mixture of any one or more.

[0056] In some embodiments, the polyethylene glycol group is selected from a functional group with a single molecular weight or a mixed functional group with multiple molecular weights.

[0057] In some embodiments, s is any one of 0, 1, 2, 3, 4, 5, and 6 or the range between any two of them.

[0058] In some embodiments, p is any one of 1, 2, 3, 4, 5, and 6 or the range between any two of them.

[0059] In some embodiments, q ranges from any one of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, and 30 to any range between any two of them.

[0060] In some embodiments, the molar ratio of Compound 1 to Compound 2 is 1:(0.1 - 40), and specifically can be in the range between any one of 1:0.1, 1:0.5, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, and 1:40 to any range between any two of them.

[0061] On the other hand, the present invention provides a method for preparing the aforementioned polymer, which includes carrying out a polymerization reaction on the aforementioned Compound 1 and Compound 2.

[0062] In some embodiments, the preparation method includes: adding an initiator when carrying out the polymerization reaction.

[0063] In some embodiments, the addition amount of the initiator is: 0.1 - 100 μM. Optionally, the addition amount of the initiator can be in the range between any one of 0.1, 0.5, 1, 2, 4, 6, 8, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, and 100 μM to any range between any two of them.

[0064] In some embodiments, the initiator includes any one or more of: tert-butyl perbenzoate, diisopropyl peroxydicarbonate, azobisisobutyronitrile, 1-hydroxy-cyclohexyl-phenyl ketone, 2-hydroxy-2-methyl-1-phenylpropanone, 1-hydroxycyclohexylbenzophenone, 2,4,6-trimethylbenzoyl diphenylphosphine oxide, and benzophenone.

[0065] In some embodiments, after adding the initiator, the preparation method further includes: carrying out thermal initiation or UV irradiation initiation when carrying out the polymerization reaction.

[0066] In some embodiments, the temperature of the thermal initiation is 50 - 150 °C, and specifically can be in the range between any one of 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, and 150 °C to any range between any two of them.

[0067] In some embodiments, before carrying out the polymerization reaction, the preparation method includes: dissolving the Compound 1 and the Compound 2 in an organic solvent.

[0068] In some embodiments, the organic solvent includes any one or a combination of more than one of: dichloromethane, chloroform, tetrahydrofuran, ethyl acetate, N,N-dimethylformamide, N-methylpyrrolidone, toluene, and benzene.

[0069] In some embodiments, the preparation method further includes: purifying the reaction solution after the polymerization reaction to obtain the final polymer.

[0070] In some embodiments, the purification includes any one or more steps of: filtration, centrifugation, washing, and drying.

[0071] In some embodiments, the detergent used for the washing includes any one or more of: tetrahydrofuran, benzene, toluene, chloroform, ethyl acetate, and anhydrous diethyl ether.

[0072] On the other hand, the present invention provides the use of the aforementioned polymer as a surfactant in the preparation of a surfactant-containing product.

[0073] On the other hand, the present invention provides a surfactant-containing product, wherein the surfactant includes the polymer described in any of the foregoing embodiments.

[0074] In some embodiments, the surfactant-containing product includes any one of: an oil phase material, a droplet, a reagent, a kit, and a chip;

[0075] In some embodiments, the oil phase material includes: an oil phase and the aforementioned polymer, and the mass ratio of the polymer to the oil phase is 0.01 to 0.1:1, specifically, it can be any one or the range between any two of 0.01:1, 0.02:1, 0.03:1, 0.04:1, 0.05:1, 0.06:1, 0.07:1, 0.08:1, 0.09:1, and 0.1:1.

[0076] In some embodiments, the oil phase material includes any one or a combination of more than one of: mineral oil, HFE 7500, HFE 7100, and FC-40.

[0077] In addition, the present invention also provides the use of the aforementioned polymer as a surfactant or the aforementioned surfactant-containing product in droplet microfluidics technology.

[0078] The present invention has no special limitation on droplet microfluidics technology. As long as the aforementioned polymer is used as a surfactant in the oil phase material to stabilize the droplet, it falls within the protection scope of this application.

[0079] In some embodiments, the applications of the droplet microfluidics technology may include: droplet microreactors (droplet microreactors utilize microfluidics technology to generate tiny droplets, enabling efficient control of chemical reactions and being applicable to various chemical reaction processes), drug transport and release (droplet microfluidics technology can be used in drug controlled-release systems to achieve precise drug delivery and release), single-cell encapsulation analysis (through droplet microfluidics technology, single cells can be encapsulated and analyzed, which is applied to biological research), droplet-based digital PCR (droplet microfluidics technology is widely used in digital PCR, improving the accuracy and efficiency of PCR), tissue engineering (droplet microfluidics technology can be used for cell culture and tissue construction in tissue engineering), diagnostic imaging (in medical diagnosis, droplet microfluidics technology can be used for the detection and imaging of biomarkers), particulate material synthesis, catalyst preparation, etc.

[0080] In some embodiments, the application in the droplet microfluidics technology is for non-disease diagnosis and / or treatment purposes.

[0081] The features and properties of the present invention will be further described in detail below in conjunction with embodiments.

[0082] Example 1

[0083] A preparation method of a surfactant, which comprises the following steps.

[0084] 1. Take 0.6 g of Compound 1 (polyethylene glycol diacrylate, PEGDA) and 2.5 g of Compound 2 (hexafluorobutyl methacrylate, MMHFB) and dissolve them in 8.4 g of dichloromethane;

[0085] The structural formula of PEGDA is:

[0086] The structural formula of MMHFB is:

[0087] 2. Add 19 mg of PBB (tert-butyl peroxybenzoate) as a thermal initiator and stir to dissolve with a magnetic rotor.

[0088] 3. React at 65 °C for 15 min while stirring with a magnetic stirrer;

[0089] 4. Transfer the reaction solution to a filtering funnel with filter paper for filtration.

[0090] 5. Add 1 mL of benzene to dissolve the polymer and elute the solvent therein;

[0091] 6. Add 1 mL of THF (tetrahydrofuran) to dissolve the polymer and elute the solvent therein;

[0092] 7. Repeat steps (5) and (6) and filter once;

[0093] 8. Vacuum dry and weigh to obtain the molecular brush fluorosurfactant powder.

[0094] 9. Weigh 0.1 g of the molecular brush fluorosurfactant powder, dissolve it in 2 g of mineral oil, shake well, and disperse it ultrasonically to obtain a 5% (w / w) molecular brush fluorosurfactant.

[0095] The infrared spectra and NMR spectra of PEGDA, MMHFB, and the product of their polymerization (PEGDA-MMHFB) are shown in Figures 2 - 7 , and the results of detecting CMC based on the DLS method and the dynamic surface tension of PEGDA-MMHFB are shown in Figure 8 .

[0096] The cost price of the surfactant prepared in Example 1 is 0.23 yuan / mL, as shown in Table 1 specifically.

[0097] Table 1 Price

[0098] Reagent Name Unit Price per ml of Synthetic Material (yuan) PEGDA 0.006 MMHFB 0.061 Dichloromethane 0.006 PBB 0.006 Benzene 0.01 THF 0.01 Mineral Oil 0.13

[0099] Example 2 has strong controllability of droplet size

[0100] Based on the surfactant provided in Example 1, 6 experimental groups were set up to prepare droplets. The processes of preparing droplets in the 6 experimental groups were generally the same, except that the water-oil flow rate ratio and the chip size were different.

[0101] The steps for preparing droplets are as follows:

[0102] (1) Prepare the continuous phase for microfluidic droplet generation

[0103] Weigh 0.2 g of the molecular brush fluorosurfactant powder, dissolve it in 4 g of mineral oil, shake well, and disperse it ultrasonically to prepare an oil phase of the surfactant with a mass concentration of 5% as the continuous phase for droplet generation.

[0104] (2) Generation of microfluidic droplets

[0105] Using water as the dispersed phase and the molecular brush fluorosurfactant (the surfactant prepared in Example 1) as the continuous phase, the generated droplets showed a high degree of uniformity. By adjusting the water-oil flow rate ratio to 1:1, 5:1, and 10:1, this example successfully generated uniform droplets with sizes ranging from 50 to 100 μm. Using microfluidic chips with cross-connection sizes of 20 μm, 50 μm, and 80 μm, uniform droplets with diameters of 25 μm, 70 μm, and 95 μm were successfully generated.

[0106] The results are shown in Figure 9, As can be seen from the results, the size of the prepared droplets can be adjusted by adjusting the water-oil flow rate ratio and the chip size, and the sizes of the prepared droplets are uniform.

[0107] Example 3 has strong versatility

[0108] Based on the surfactant provided in Example 1, 4 experimental groups were set up to prepare droplets. The processes of preparing droplets in the 4 experimental groups were generally the same, except that different aqueous phases were used.

[0109] The steps for preparing droplets are as follows:

[0110] (1) Prepare the continuous phase for generating microfluidic droplets

[0111] Weigh 0.2 g of the molecular brush fluorinated surfactant powder and dissolve it in 4 g of mineral oil. Shake well and disperse ultrasonically to prepare an oil phase of the surfactant with a mass concentration of 5%, which is used as the continuous phase for droplet generation.

[0112] (2) Generation of microfluidic droplets

[0113] Respectively draw a certain amount of the dispersed phase (H 2 O, phosphate buffered saline (PBS), Luria-Bertani (LB) medium, and yeast extract peptone dextrose (YPD) medium) and the continuous phase (oil containing the molecular brush fluorinated surfactant) into two disposable syringes. Push the dispersed phase and the continuous phase liquids into the microfluidic chip at a flow rate ratio of 1:3 by using a micro-injection pump, and observe the generation of droplets in real time under a microscope. Collect the droplets at the chip outlet.

[0114] (3) Measurement of droplet size

[0115] Measure the droplet sizes of different aqueous phases placed at room temperature for different times on the Image J software, and draw a statistical chart with the origin software.

[0116] The results are shown in Figure 10 , As can be seen from the results, the droplets of different aqueous phases prepared by the surfactant prepared in the embodiment of the present invention have uniformity and good stability.

[0117] Example 4 has good stability

[0118] Perform a stability test on the surfactant prepared in Example 1. The test methods include:

[0119] 1. First, the ability of droplets generated by molecular brush surfactants to be stored for a long time was verified. Weigh 0.2 g of the molecular brush fluorosurfactant powder and dissolve it in 4 g of mineral oil. Shake well and disperse ultrasonically to prepare an oil phase of surfactant with a mass concentration of 5%, which serves as the continuous phase for droplet generation. Draw a certain amount of the dispersed phase (H 2 O) and the continuous phase (oil containing the molecular brush fluorosurfactant) into two disposable syringes respectively. Push the dispersed phase and the continuous phase liquids into the microfluidic chip at a flow rate ratio of 1:3 using a micro-injection pump, and observe the droplet generation in real time under a microscope. Collect the droplets at the chip outlet. Collect 500 μL of droplets and place them at room temperature for 7 days. By statistically analyzing the droplet diameter distribution at 0 day and 7 days, it is found that the droplets stored for a long time have good uniformity, and there is no significant droplet fusion and rupture.

[0120] 2. The collected droplets were subjected to PCR thermal cycling. The cycling was carried out 30 times under the conditions of 95° (30 s), 60° (30 s), and 72° (30 s). After the thermal cycling, compared with before cycling, there was no large-area fusion of the droplets, and the droplets still maintained their integrity. By statistically analyzing the droplet diameter distribution, it can be seen that there is no obvious change in the droplet diameter before and after the thermal cycling.

[0121] The results are shown in Figure 11 . It can be seen from the results that the droplets prepared by the surfactant prepared in the embodiments of the present invention can maintain their droplet morphology and have good thermal stability even after being placed for a long time.

[0122] Example 5 has good biocompatibility

[0123] Use the surfactant prepared in Example 1 to prepare droplets. The steps for preparing the droplets include: weigh 0.2 g of the molecular brush fluorosurfactant powder and dissolve it in 4 g of mineral oil. Shake well and disperse ultrasonically to prepare an oil phase of surfactant with a mass concentration of 5%, which serves as the continuous phase for droplet generation. Draw a certain amount of the dispersed phase (E. coli / yeast cells) and the continuous phase (oil containing the molecular brush fluorosurfactant) into two disposable syringes respectively. Push the dispersed phase and the continuous phase liquids into the microfluidic chip at a flow rate ratio of 1:3 using a micro-injection pump, and observe the droplet generation in real time under a microscope. Collect the droplets at the chip outlet.

[0124] Encapsulate 0.6 E. coli. / droplet (~65 μm, about 4.2×10 6 CFU / mL) into single droplets for single-bacterium proliferation. Perform fluorescence imaging at 0, 12, 24, 36, and 48 hours respectively, and reflect the bacterial proliferation in the droplets by the size of the fluorescence area. The results are shown in Figure 12 A in.

[0125] Single-cell proliferation was carried out by encapsulating 0.6 Yeast / droplet (~65 μm, approximately 4.2×10 6 Yeast / mL) into individual droplets, and imaging was performed at 0, 3, 6, 9, 12, and 24 hours. The growth of individual yeast cells in the droplets caused the diameter of the droplets containing yeast cells to gradually decrease. The results are shown in Figure 12 B in

[0126] It can be clearly seen that the surfactant prepared in the examples of the present invention has good biocompatibility.

[0127] When the molar ratio of compound 1 to compound 2 in Example 6 was 1:1, the thermal stability of the droplets was the best.

[0128] To evaluate the effect of the polymerization ratio of the two compounds on the thermal stability of the droplets. Compound 1 (PEGDA) and compound 2 (MMHFB) were mixed at a molar concentration of 1 μM of compound 1 and a molar ratio of 1:0.1, 1:1, and 1:5 of compound 1 to compound 1 to prepare 3 kinds of molecular brush fluorinated surfactants. Weigh 0.2 g of different kinds of molecular brush fluorinated surfactant powders, dissolve them in 4 g of mineral oil, shake and mix well, and disperse them ultrasonically to prepare an oil phase of a surfactant with a mass concentration of 5%, which is used as the continuous phase for droplet generation. A certain amount of the dispersed phase (H 2 O) and 3 different continuous phases (oil containing molecular brush fluorinated surfactant) were respectively drawn into a disposable syringe, and the dispersed phase and the continuous phase liquids were pushed into the microfluidic chip at a flow rate ratio of 1:3 by using a micro-injection pump, and the generation of droplets was observed in real time under a microscope. Droplets generated by 3 kinds of molecular brush fluorinated surfactants were collected at the chip outlet. 500 μL of droplets were collected for PCR thermal cycling, and cycled 30 times under the conditions of 95° for 30 s, 60° for 30 s, and 72° for 30 s. Compared with before cycling, large-area fusion occurred when the molar ratio of compound 1 to compound 2 was 1:0.1 and 1:5. However, the diameter of the droplets generated when the molar ratio of compound 1 to compound 2 was 1:1 did not change significantly.

[0129] The results are shown in Figure 13 . From the results, it can be seen that the droplets prepared with the surfactant prepared in the examples of the present invention have good thermal stability, and the droplets generated when the molar ratio of compound 1 to compound 2 is 1:1 have the best stability.

[0130] Compared with other fluorinated surfactants in Example 7, the droplets also have good stability.

[0131] To evaluate the effect of comparing the droplet stability with other fluorosurfactants, the ability of droplets generated by three fluorosurfactants to be stored for a long time was verified. Weigh 0.2 g of fluorosurfactants PEG–PFPE (commercially available), PEG–PFPE 2 (commercially available), and PEGDA-MMHFB powder, dissolve them in 4 g of mineral oil, shake and mix well, and disperse them by ultrasound to prepare an oil phase of surfactant with a mass concentration of 5%, which serves as the continuous phase for droplet generation. Draw a certain amount of dispersed phase (H 2 O) and continuous phase (oil containing three fluorosurfactants) with a disposable syringe, and push the dispersed phase and continuous phase liquids into the microfluidic chip at a flow rate ratio of 1:3 by using a microinjection pump, and observe the droplet generation situation in real time under a microscope, and collect droplets at the chip outlet. Collect 500 μL of droplets and place them at room temperature for 7 days. By statistically analyzing the droplet diameter distribution at 0 day and 7 days, it is found that the uniformity of the droplets using PEGDA-MMHFB is the best after long-term placement, and there is no droplet fusion and rupture.

[0132] The results are shown in Figure 14 , and it can be seen from the results that the droplets prepared by the surfactant prepared in the embodiment of the present invention have more excellent stability than other fluorosurfactants.

[0133] Test on the stability and biocompatibility of the surfactant obtained by copolymerizing different compound 1 with MMHFB in Example 8

[0134] To verify the effect of the copolymer formed by compound 1 with other structures and MMHFB, the applicant synthesized the following compounds 1-1 to 1-6, and copolymerized them with MMHFB under the same conditions as PEGDA to obtain copolymers. Using these copolymers as surfactants, the same stability and biocompatibility tests as in Examples 4 and 5 were carried out. The results showed that although the stability and biocompatibility were slightly weaker than those of PEGDA, they were still excellent, and better than those of commercially available PEG–PFPE 2 .

[0135] Compound 1-1 is a compound in which R 1 in Formula 1 is NH and the remaining structure is the same as that of PEGDA;

[0136] Compound 1-2 is a compound in which R 2 in Formula 1 is CH 3 and the remaining structure is the same as that of PEGDA;

[0137] Compound 1-3 is a compound in which R 2 in Formula 1 is -OH and the remaining structure is the same as that of PEGDA;

[0138] Compounds 1-4 are compounds in which r is 2 in Formula 1 and the rest of the structure is the same as that of PEGDA;

[0139] Compounds 1-5 are compounds in which r is 20 in Formula 1 and the rest of the structure is the same as that of PEGDA;

[0140] Compounds 1-6 are compounds in which s is 6 in Formula 1 and the rest of the structure is the same as that of PEGDA.

[0141] Test on the stability and biocompatibility of surfactants obtained by copolymerizing different Compound 2 with PEGDA in Example 9.

[0142] In order to verify the effects of copolymers formed by Compound 2 with other structures and PEGDA, the applicant synthesized the following Compounds 2-1 to 2-6, copolymerized them with PEGDA under the same conditions as MMHFB to obtain copolymers, and used these copolymers as surfactants to conduct the same stability and biocompatibility tests as in Examples 4 and 5 above. It was found that although the stability and biocompatibility were slightly weaker than those of MMHFB, they were still excellent, and better than those of commercially available PEG–PFPE 2 .

[0143] Compound 2-1 is a compound in which R 3 is NH in Formula 2 and the rest of the structure is the same as that of MMHFB (i.e., replacing -O- in hexafluorobutyl methacrylate with -NH-);

[0144] Compound 2-2 is a compound in which R 4 is CF 2 and p is 1, q is 5, and the rest of the structure is the same as that of MMHFB;

[0145] Compound 2-3 is a compound in which R 4 is CHF and p is 1, q is 5, and the rest of the structure is the same as that of MMHFB;

[0146] Compound 2-4 is a compound in which R 4 is CF 2 and p is 6, q is 2, and the rest of the structure is the same as that of MMHFB;

[0147] Compound 2-5 is a compound in which R 4 is CHF and p is 6, q is 2, and the rest of the structure is the same as that of MMHFB;

[0148] Compound 2-6 is a compound in which R 4 is CF 2 and p is 3, q is 2, and the rest of the structure is the same as that of MMHFB.

[0149] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A polymer, characterized in that Obtained by copolymerization of compound 1 and compound 2, The structural formula of the compound 1 is shown in Formula 1; Formula 1: Wherein, R1 is O or NH, R2 is selected from any one of: an alkyl group having 1 to 5 carbon atoms, a hydroxyl group, and CH2=CH-C(=O)-, r is an integer of 2 to 500, and s is an integer of 0 to 6; The structural formula of the compound 2 is shown in Formula 2; Formula 2: Wherein, R3 is O or NH, R4 is CF2, CHF or CF2CHF, p is an integer of 1 to 6, and q is an integer of 1 to 30.

2. The polymer according to claim 1, characterized in that The molar ratio of the compound 1 to the compound 2 is 1:(0.1-40); Optionally, the molar ratio is 1:(0.1-5); Optionally, the molecular weight of the polyethylene glycol group in Formula 1 is 50 to 10,000; Optionally, the polyethylene glycol group is selected from a functional group of a single molecular weight or a mixed functional group of multiple molecular weights.

3. The method for preparing a polymer according to claim 1 or 2, characterized in that: The method comprises the steps of polymerizing compound 1 and compound 2.

4. The preparation method according to claim 3, characterized in that: adding an initiator during the polymerization reaction; Optionally, the added amount of the initiator is: 0.1 to 100 μM; Optionally, the initiator includes: any one or more combinations of tert-butyl perbenzoate, diisopropyl peroxydicarbonate, azobisisobutyronitrile, 1-hydroxy-cyclohexyl-phenyl ketone, 2-hydroxy-2-methyl-1-phenylacetone, 1-hydroxycyclohexyl benzophenone, 2,4,6-trimethylbenzoyldiphenylphosphine oxide and benzophenone; Optionally, after adding the initiator, the preparation method further comprises: performing thermal initiation or UV irradiation photoinitiation during the polymerization reaction; Optionally, the thermal initiation temperature is 50-150°C.

5. The preparation method according to claim 3, characterized in that: Before performing the polymerization reaction, dissolving the compound 1 and the compound 2 in an organic solvent; Optionally, the organic solvent includes any one or more combinations of dichloromethane, chloroform, tetrahydrofuran, ethyl acetate, N,N-dimethylformamide, N-methylpyrrolidone, toluene and benzene.

6. The preparation method according to any one of claims 3 to 5, characterized in that: The preparation method also includes a step of purifying the reaction solution after the polymerization reaction to obtain the final polymer; Optionally, the purification comprises: any one or more steps of filtration, centrifugation, washing and drying; Optionally, the washing agent used in the washing includes any one or more of tetrahydrofuran, benzene, toluene, chloroform, ethyl acetate and anhydrous ether.

7. Use of the polymer according to claim 1 or 2 as a surfactant in the preparation of a product containing a surfactant.

8. A product containing a surfactant, characterized in that: The surfactant comprises the polymer according to claim 1 or 2.

9. The surfactant-containing product according to claim 8, characterized in that The surfactant-containing product includes any one of an oil phase material, a droplet, a reagent, a kit and a chip.

10. Use of the polymer according to claim 1 or 2 as a surfactant or the product containing a surfactant according to claim 8 or 9 in droplet microfluidics technology.

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