A low-slip-rate adhesive glass slide and its preparation method

Abstract

The present invention relates to the technical field of glass slides, and discloses a low-slip glass slide and a preparation method thereof; the preparation method includes the following steps: soaking a thermally-alkali-activated glass slide in a poly-L-lysine solution, removing and washing with deionized water to obtain a layer of poly-L-lysine film; dropping a dialdehyde β-cyclodextrin-modified oxidized sodium alginate solution onto the poly-L-lysine film, spin-coating, washing, and drying to obtain a preliminary low-slip glass slide; dropping a poly-L-lysine solution onto the preliminary low-slip glass slide, spin-coating, washing, then dropping a dialdehyde β-cyclodextrin-modified oxidized sodium alginate solution, spin-coating, washing. Repeating the above steps of dropping the poly-L-lysine solution and the dialdehyde β-cyclodextrin-modified oxidized sodium alginate solution for one spin-coating, and repeating the spin-coating step n times to prepare a low-slip glass slide with low detachment rate.

Description

Technical Field

[0001] The invention relates to the technical field of glass slides, in particular to an adhesive glass slide with a low detachment rate and a preparation method thereof. Background Art

[0002] In the biomedical field, glass slides are widely used in cell culture, cell observation, and biological analysis. However, traditional substrate adhesion materials have a series of problems in the application process, which limits their wide application in high-performance biomedical applications. Traditional substrate adhesion and hydrophilic modification materials represented by dopamine have the problem of oxidative self-polymerization reaction, resulting in the formation of melanin polymers, which significantly reduces the light transmittance of the dopamine deposition film after oxidative polymerization, limiting its effect in applications such as microscopic observation.

[0003] In addition, although the hydrophilic coating formed by directly immersing the glass slide in poly-L-lysine can achieve a certain degree of hydrophilicity, its own substrate adhesion performance is relatively poor, which makes it easy to peel off in actual applications, affecting the long-term stability and reliability of the glass slide.

[0004] When chitosan is used as a component in the preparation of multilayer composite membrane coatings, chitosan is prone to swelling, which reduces its modulus and has a negative impact on cell adhesion, resulting in a decrease in the number of cell adhesions, which restricts its applicability in biomedical applications such as cell culture.

[0005] Therefore, in order to overcome the problems existing in traditional substrate adhesion materials, improve the light transmittance and substrate adhesion performance of the slide, and ensure good cell adhesion properties in biomedical applications such as cell culture, it is urgently necessary to provide a low detachment rate, high-performance adhesion slide and its preparation method. Summary of the invention

[0006] The object of the present invention is to provide an adhesive glass slide with low detachment rate and a preparation method thereof, so as to solve the problems raised in the above background technology.

[0007] In order to solve the above technical problems, the present invention provides the following technical solutions:

[0008] A method for preparing an adhesive glass slide with a low detachment rate comprises the following steps:

[0009] S1: soaking a hot alkali activated glass slide in a poly-L-lysine solution, taking it out and washing it with deionized water to obtain a layer of poly-L-lysine film; dropping a dialdehyde β-cyclodextrin modified oxidized sodium alginate solution on the poly-L-lysine film, spin coating, washing, and drying to obtain a prepared low-detachment rate glass slide;

[0010] S2: Add the poly-L-lysine solution dropwise onto the prepared low detachment rate glass slide, spin-coat, wash, then add the dialdehyde β-cyclodextrin modified oxidized sodium alginate solution dropwise, spin-coat, wash, repeat the above poly-L-lysine solution and dialdehyde β-cyclodextrin modified oxidized sodium alginate solution as one spin coating, repeat the spin coating steps n times to prepare a low detachment rate adhesive glass slide.

[0011] Furthermore, the method for preparing the hot alkali activated glass slide comprises the following steps:

[0012] Step (1): After washing the slide glass with deionized water, soak it in a potassium permanganate solution for 4-8 hours, take it out and wash it with deionized water, and finally ultrasonically clean it with deionized water, and dry it at 80-85° C. to obtain a pretreated slide glass;

[0013] Step (2): using the pretreated glass slide as a substrate, placing a patterned grating template, and magnetron sputtering to obtain a titanium thin film glass slide;

[0014] Step (3): placing the titanium film slide in a sodium hydroxide solution, activating, washing, nitrogen purging, and drying to obtain a hot alkali activated slide.

[0015] Furthermore, in step (2), the magnetron sputtering target is a Ti target, the magnetron sputtering time is 7-9 min, and the vacuum degree is 5.5×10 -4 -6.0×10 -4 Pa, argon flow rate is 20-30sccm, chamber pressure is 0.15-0.18Pa, and sputtering current is 0.16-0.19A.

[0016] Furthermore, in step (3), the concentration of the sodium hydroxide solution is 0.1-0.2 mol / L; the activation temperature is 45-50° C., and the activation time is 25-30 min.

[0017] Furthermore, the method for preparing the dialdehyde β-cyclodextrin modified oxidized sodium alginate solution comprises the following steps:

[0018] Place sodium alginate in a reaction container, add a buffer solution and stir evenly, add a cross-linking aid and stir to activate, add mono-(6-amino-6-deoxy)-β-cyclodextrin solution and stir evenly, react at room temperature for 22-26 hours, purify, and freeze-dry to obtain cyclodextrin-modified sodium alginate; add sodium periodate solution to the cyclodextrin-modified sodium alginate solution, stir evenly in a light-proof condition, react for 20-24 hours, add ethylene glycol to terminate the reaction, purify, and obtain a dialdehyde β-cyclodextrin-modified oxidized sodium alginate solution.

[0019] Furthermore, the buffer solution is a phosphate buffer solution with a pH of 7.2-7.3; the cross-linking aid is a mixture of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide in a molar ratio of 1:1; the solvent in the cross-linking aid and mono-(6-amino-6-deoxy)-β-cyclodextrin solution is a buffer solution; the molar ratio of sodium alginate:cross-linking aid is (1-1.2):(6-6.4); the molar ratio of sodium alginate:mono-(6-amino-6-deoxy)-β-cyclodextrin is (1-1.2):(2-2.4).

[0020] Furthermore, the concentration of the cyclodextrin modified sodium alginate solution is 1.2-1.5 wt %.

[0021] Furthermore, in step S1, the concentration of the poly-L-lysine solution is 1-2 mg / mL, and the concentration of the dialdehyde β-cyclodextrin modified oxidized sodium alginate solution is 1-2 mg / mL.

[0022] Furthermore, in step S2, the concentration of the poly-L-lysine solution is 0.5-1 mg / mL, and the concentration of the dialdehyde β-cyclodextrin modified oxidized sodium alginate solution is 1-2 mg / mL; the spin coating is repeated n times n=8-12 times, and the coating thickness of one spin coating is 0.25-0.3 μm.

[0023] Furthermore, the molar ratio of sodium periodate in the sodium periodate solution to cyclodextrin-modified sodium alginate in the cyclodextrin-modified sodium alginate solution is 0.5:1.

[0024] Furthermore, the molar ratio of ethylene glycol to sodium periodate is 1:1.

[0025] Compared with the prior art, the beneficial effects achieved by the present invention are:

[0026] Traditional dopamine is used as a traditional substrate adhesion and hydrophilic modification material, and there is an oxidative self-polymerization reaction and subsequent reaction that will be converted into a melanin polymer, resulting in the problem that the color of the polydopamine deposited film after oxidative polymerization is darker and the light transmittance is low; and the hydrophilic coating formed by directly immersing the glass slide in poly-L-polylysine has poor substrate adhesion performance of the coating itself; in order to solve the above technical problems, the present invention sets a pattern grating template on the surface of the glass slide, controls the parameters of magnetron sputtering by magnetron sputtering, especially controls the magnetron sputtering time, and uses a Ti target as a target material to magnetron sputter a layer of nano-scale titanium film on the surface of an ordinary glass slide, while meeting the light transmittance requirements of the glass slide and reserving a sample area, and then anionizes the titanium film surface by hot alkali activation, and uses poly-L-lysine itself to provide chiral polycations to generate strong electrostatic interaction with the anionized titanium film surface, thereby greatly improving the substrate adhesion performance of the coating, while avoiding the problem of low light transmittance caused by melanin polymers brought by polydopamine. At the same time, the particles of the magnetron sputtered titanium film itself have the characteristic of photo-induced hydrophilicity, which greatly reduces the wetting angle to water and further improves the hydrophilicity of the glass slide.

[0027] The invention realizes an acylation reaction of sodium alginate under the activation of EDC / NHS, successfully grafts mono-(6-amino-6-deoxy)-β-cyclodextrin on the sodium alginate, and then simultaneously oxidizes the sodium alginate and β-cyclodextrin into dialdehyde oxidized sodium alginate and dialdehyde β-cyclodextrin through sodium periodate oxidation, so as to prepare dialdehyde β-cyclodextrin modified oxidized sodium alginate; the aldehyde group of the dialdehyde β-cyclodextrin modified oxidized sodium alginate and the amino group on the surface of poly-L-lysine are used to generate a Schiff base reaction through dehydration condensation to form a dynamic imine bond, and the carboxyl anion of the poly-L-lysine and the amino cation of the dialdehyde β-cyclodextrin modified oxidized sodium alginate have an electrostatic adsorption effect, so as to form a layer-by-layer self-assembled composite network structure. The Schiff base combined coating has excellent bioadhesion, further improving the substrate adhesion and cell adhesion performance of the glass slide; it solves the problem that when traditional chitosan is used as a cell adhesion polyelectrolyte to prepare a multilayer composite membrane coating, it will swell in the solution, resulting in a decrease in modulus, affecting cell adhesion and reducing the number of cell adhesion. DETAILED DESCRIPTION

[0028] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0029] In the following examples, the glass slide is a smooth glass slide purchased from Jiangsu Huida Medical Instrument Co., Ltd.; the Ti target specification is φ100mm×6mm, purchased from Zhongnuo New Materials (Beijing) Technology Co., Ltd.; the sodium alginate specification is Mw=60000, purchased from Sinopharm Chemical Reagent Co., Ltd.; the molecular weight of poly-L-lysine is 150000-300000, and the molecular weight of poly-D-lysine is 150000-300000, purchased from Sigma-Aldrich; mono-(6-amino-6-deoxy)-β-cyclodextrin is purchased from Shanghai MacLean Biochemical Technology Co., Ltd.; the remaining raw materials are commercially available.

[0030] Example 1: A method for preparing an adhesive glass slide with a low detachment rate: S1: After washing the glass slide with deionized water, soak it in a potassium permanganate solution for 6 hours, take it out and wash it with deionized water, and finally ultrasonically clean it with deionized water, and dry it at 80°C to obtain a pretreated glass slide;

[0031] S2: The pretreated glass slide was used as the substrate, and the pattern grating template was placed, and magnetron sputtering was performed to obtain a titanium thin film glass slide; wherein the magnetron sputtering target material was Ti target, the magnetron sputtering time was 7 min, and the vacuum degree was 5.5×10 -4 Pa, argon flow rate is 20sccm, chamber pressure is 0.15Pa, sputtering current is 0.16A

[0032] S3: placing the titanium film slide in a 0.2 mol / L sodium hydroxide solution, activating at 50°C for 30 min, washing, purging with nitrogen, and drying to obtain a hot alkali activated slide;

[0033] S4: Place 1mmol sodium alginate in a reaction container, add pH 7.2 phosphate buffer solution, stir evenly, add 3mmol 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and 3mmol N-hydroxysuccinimide, stir to activate, add 2mmol mono-(6-amino-6-deoxy)-β-cyclodextrin solution, stir evenly, react at room temperature for 24h, purify, freeze-dry, and obtain cyclodextrin-modified sodium alginate; add sodium periodate solution to 1.2wt% cyclodextrin-modified sodium alginate solution, stir evenly in a dark environment, react for 24h, add ethylene glycol to terminate the reaction, purify, and obtain dialdehyde β-cyclodextrin-modified oxidized sodium alginate solution; wherein the molar ratio of sodium periodate in the sodium periodate solution: cyclodextrin-modified sodium alginate: ethylene glycol in the cyclodextrin-modified sodium alginate solution is 0.5:1:0.5;

[0034] S5: soaking the hot alkali activated glass slide in a 1 mg / mL poly-L-lysine solution, taking it out and washing it with deionized water to obtain a layer of poly-L-lysine film; dropping 1 mg / mL dialdehyde β-cyclodextrin modified oxidized sodium alginate solution on the poly-L-lysine film, spin coating, washing, and drying to obtain a prepared low-detachment rate glass slide; wherein the thickness of the dialdehyde β-cyclodextrin modified oxidized sodium alginate layer is 0.25 μm;

[0035] S6: Add 0.5 mg / mL poly-L-lysine solution on the prepared low detachment rate slide glass, spin coat, wash, then add 1 mg / mL dialdehyde β-cyclodextrin modified oxidized sodium alginate solution, spin coat, wash, repeat the above poly-L-lysine solution and dialdehyde β-cyclodextrin modified oxidized sodium alginate solution for one spin coating, the coating thickness of one spin coating is 0.25 μm; repeat the spin coating steps 8 times to prepare a low detachment rate adhesive slide glass.

[0036] Example 2: A method for preparing an adhesive glass slide with a low detachment rate: S1: After washing the glass slide with deionized water, soak it in a potassium permanganate solution for 6 hours, take it out and wash it with deionized water, and finally ultrasonically clean it with deionized water, and dry it at 80°C to obtain a pretreated glass slide;

[0037] S2: The pretreated glass slide was used as the substrate, and the pattern grating template was placed, and magnetron sputtering was performed to obtain a titanium thin film glass slide; wherein the magnetron sputtering target material was Ti target, the magnetron sputtering time was 8 min, and the vacuum degree was 5.5×10 -4 Pa, argon flow rate is 20sccm, chamber pressure is 0.15Pa, sputtering current is 0.16A

[0038] S3: placing the titanium film slide in a 0.2 mol / L sodium hydroxide solution, activating at 50°C for 30 min, washing, purging with nitrogen, and drying to obtain a hot alkali activated slide;

[0039] S4: 1mmol sodium alginate is placed in a reaction container, pH 7.2 phosphate buffer solution is added, stirred evenly, 3mmol 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and 3mmol N-hydroxysuccinimide are added, stirred for activation, 2mmol mono-(6-amino-6-deoxy)-β-cyclodextrin solution is added, stirred evenly, reacted at room temperature for 24h, purified, freeze-dried, and cyclodextrin-modified sodium alginate is obtained; sodium periodate solution is added to 1.2wt% cyclodextrin-modified sodium alginate solution, stirred evenly in a light-proof condition, reacted for 24h, ethylene glycol is added to terminate the reaction, purified, and dialdehyde β-cyclodextrin-modified oxidized sodium alginate solution is obtained; wherein the molar ratio of sodium periodate in the sodium periodate solution: cyclodextrin-modified sodium alginate: ethylene glycol in the cyclodextrin-modified sodium alginate solution is 0.5:1:0.5;

[0040] S5: soaking the hot alkali activated glass slide in a 1 mg / mL poly-L-lysine solution, taking it out and washing it with deionized water to obtain a layer of poly-L-lysine film; dropping 1 mg / mL dialdehyde β-cyclodextrin modified oxidized sodium alginate solution on the poly-L-lysine film, spin coating, washing, and drying to obtain a prepared low-detachment rate glass slide; wherein the thickness of the dialdehyde β-cyclodextrin modified oxidized sodium alginate layer is 0.25 μm;

[0041] S6: Add 0.5 mg / mL poly-L-lysine solution on the prepared low detachment rate slide glass, spin coat, wash, then add 1 mg / mL dialdehyde β-cyclodextrin modified oxidized sodium alginate solution, spin coat, wash, repeat the above poly-L-lysine solution and dialdehyde β-cyclodextrin modified oxidized sodium alginate solution for one spin coating, the coating thickness of one spin coating is 0.25 μm; repeat the spin coating steps 8 times to prepare a low detachment rate adhesive slide glass.

[0042] Example 3: A method for preparing an adhesive glass slide with a low detachment rate: S1: After washing the glass slide with deionized water, soak it in a potassium permanganate solution for 6 hours, take it out and wash it with deionized water, and finally ultrasonically clean it with deionized water, and dry it at 80°C to obtain a pretreated glass slide;

[0043] S2: The pretreated glass slide was used as the substrate, and the pattern grating template was placed, and magnetron sputtering was performed to obtain a titanium thin film glass slide; wherein the magnetron sputtering target material was Ti target, the magnetron sputtering time was 9 min, and the vacuum degree was 5.5×10 -4 Pa, argon flow rate is 20sccm, chamber pressure is 0.15Pa, sputtering current is 0.16A

[0044] S3: placing the titanium film slide in a 0.2 mol / L sodium hydroxide solution, activating at 50°C for 30 min, washing, purging with nitrogen, and drying to obtain a hot alkali activated slide;

[0045] S4: Place 1mmol sodium alginate in a reaction container, add pH 7.2 phosphate buffer solution, stir evenly, add 3mmol 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and 3mmol N-hydroxysuccinimide, stir to activate, add 2mmol mono-(6-amino-6-deoxy)-β-cyclodextrin solution, stir evenly, react at room temperature for 24h, purify, freeze-dry, and obtain cyclodextrin-modified sodium alginate; add sodium periodate solution to 1.2wt% cyclodextrin-modified sodium alginate solution, stir evenly in a dark environment, react for 24h, add ethylene glycol to terminate the reaction, purify, and obtain dialdehyde β-cyclodextrin-modified oxidized sodium alginate solution; wherein the molar ratio of sodium periodate in the sodium periodate solution: cyclodextrin-modified sodium alginate: ethylene glycol in the cyclodextrin-modified sodium alginate solution is 0.5:1:0.5;

[0046] S5: soaking the hot alkali activated glass slide in a 1 mg / mL poly-L-lysine solution, taking it out and washing it with deionized water to obtain a layer of poly-L-lysine film; dropping 1 mg / mL dialdehyde β-cyclodextrin modified oxidized sodium alginate solution on the poly-L-lysine film, spin coating, washing, and drying to obtain a prepared low-detachment rate glass slide; wherein the thickness of the dialdehyde β-cyclodextrin modified oxidized sodium alginate layer is 0.25 μm;

[0047] S6: Add 0.5 mg / mL poly-L-lysine solution on the prepared low detachment rate slide glass, spin coat, wash, then add 1 mg / mL dialdehyde β-cyclodextrin modified oxidized sodium alginate solution, spin coat, wash, repeat the above poly-L-lysine solution and dialdehyde β-cyclodextrin modified oxidized sodium alginate solution for one spin coating, the coating thickness of one spin coating is 0.25 μm; repeat the spin coating steps 8 times to prepare a low detachment rate adhesive slide glass.

[0048] Example 4: A method for preparing an adhesive glass slide with a low detachment rate: S1: After washing the glass slide with deionized water, soak it in a potassium permanganate solution for 6 hours, take it out and wash it with deionized water, and finally ultrasonically clean it with deionized water, and dry it at 80°C to obtain a pretreated glass slide;

[0049] S2: The pretreated glass slide was used as the substrate, and the pattern grating template was placed, and magnetron sputtering was performed to obtain a titanium thin film glass slide; wherein the magnetron sputtering target material was Ti target, the magnetron sputtering time was 9 min, and the vacuum degree was 5.5×10 -4 Pa, argon flow rate is 20sccm, chamber pressure is 0.15Pa, sputtering current is 0.16A

[0050] S3: placing the titanium film slide in a 0.2 mol / L sodium hydroxide solution, activating at 50°C for 30 min, washing, purging with nitrogen, and drying to obtain a hot alkali activated slide;

[0051] S4: Place 1mmol sodium alginate in a reaction container, add pH 7.2 phosphate buffer solution, stir evenly, add 3mmol 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and 3mmol N-hydroxysuccinimide, stir to activate, add 2mmol mono-(6-amino-6-deoxy)-β-cyclodextrin solution, stir evenly, react at room temperature for 24h, purify, freeze-dry, and obtain cyclodextrin-modified sodium alginate; add sodium periodate solution to 1.2wt% cyclodextrin-modified sodium alginate solution, stir evenly in a dark environment, react for 24h, add ethylene glycol to terminate the reaction, purify, and obtain dialdehyde β-cyclodextrin-modified oxidized sodium alginate solution; wherein the molar ratio of sodium periodate in the sodium periodate solution: cyclodextrin-modified sodium alginate: ethylene glycol in the cyclodextrin-modified sodium alginate solution is 0.5:1:0.5;

[0052] S5: soaking the hot alkali activated glass slide in a 1 mg / mL poly-L-lysine solution, taking it out and washing it with deionized water to obtain a layer of poly-L-lysine film; dropping 1 mg / mL dialdehyde β-cyclodextrin modified oxidized sodium alginate solution on the poly-L-lysine film, spin coating, washing, and drying to obtain a prepared low-detachment rate glass slide; wherein the thickness of the dialdehyde β-cyclodextrin modified oxidized sodium alginate layer is 0.25 μm;

[0053] S6: Add 0.5 mg / mL poly-L-lysine solution on the prepared low detachment rate slide glass, spin coat, wash, then add 1 mg / mL dialdehyde β-cyclodextrin modified oxidized sodium alginate solution, spin coat, wash, repeat the above poly-L-lysine solution and dialdehyde β-cyclodextrin modified oxidized sodium alginate solution for one spin coating, the coating thickness of one spin coating is 0.25 μm; repeat the spin coating step 10 times to prepare a low detachment rate adhesive slide glass.

[0054] Example 5: A method for preparing an adhesive glass slide with a low detachment rate: S1: After washing the glass slide with deionized water, soak it in a potassium permanganate solution for 6 hours, take it out and wash it with deionized water, and finally ultrasonically clean it with deionized water, and dry it at 80°C to obtain a pretreated glass slide;

[0055] S2: The pretreated glass slide was used as the substrate, and the pattern grating template was placed, and magnetron sputtering was performed to obtain a titanium thin film glass slide; wherein the magnetron sputtering target material was Ti target, the magnetron sputtering time was 9 min, and the vacuum degree was 5.5×10 -4 Pa, argon flow rate is 20sccm, chamber pressure is 0.15Pa, sputtering current is 0.16A

[0056] S3: placing the titanium film slide in a 0.2 mol / L sodium hydroxide solution, activating at 50°C for 30 min, washing, purging with nitrogen, and drying to obtain a hot alkali activated slide;

[0057] S4: Place 1mmol sodium alginate in a reaction container, add pH 7.2 phosphate buffer solution, stir evenly, add 3mmol 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and 3mmol N-hydroxysuccinimide, stir to activate, add 2mmol mono-(6-amino-6-deoxy)-β-cyclodextrin solution, stir evenly, react at room temperature for 24h, purify, freeze-dry, and obtain cyclodextrin-modified sodium alginate; add sodium periodate solution to 1.2wt% cyclodextrin-modified sodium alginate solution, stir evenly in a dark environment, react for 24h, add ethylene glycol to terminate the reaction, purify, and obtain dialdehyde β-cyclodextrin-modified oxidized sodium alginate solution; wherein the molar ratio of sodium periodate in the sodium periodate solution: cyclodextrin-modified sodium alginate: ethylene glycol in the cyclodextrin-modified sodium alginate solution is 0.5:1:0.5;

[0058] S5: soaking the hot alkali activated glass slide in a 1 mg / mL poly-L-lysine solution, taking it out and washing it with deionized water to obtain a layer of poly-L-lysine film; dropping 1 mg / mL dialdehyde β-cyclodextrin modified oxidized sodium alginate solution on the poly-L-lysine film, spin coating, washing, and drying to obtain a prepared low-detachment rate glass slide; wherein the thickness of the dialdehyde β-cyclodextrin modified oxidized sodium alginate layer is 0.25 μm;

[0059] S6: Add 0.5 mg / mL poly-L-lysine solution on the prepared low detachment rate glass slide, spin coat, wash, then add 1 mg / mL dialdehyde β-cyclodextrin modified oxidized sodium alginate solution, spin coat, wash, repeat the above poly-L-lysine solution and dialdehyde β-cyclodextrin modified oxidized sodium alginate solution for one spin coating, the coating thickness of one spin coating is 0.25 μm; repeat the spin coating step 12 times to prepare a low detachment rate adhesive glass slide.

[0060] Comparative Example 1: A method for preparing an adhesive glass slide with a low detachment rate: S1: After washing the glass slide with deionized water, soak it in a potassium permanganate solution for 6 hours, take it out and wash it with deionized water, and finally ultrasonically clean it with deionized water, and dry it at 80°C to obtain a pretreated glass slide;

[0061] S2: The pretreated glass slide was used as the substrate, and the pattern grating template was placed, and magnetron sputtering was performed to obtain a titanium thin film glass slide; wherein the magnetron sputtering target material was Ti target, the magnetron sputtering time was 7 min, and the vacuum degree was 5.5×10 -4 Pa, argon flow rate is 20sccm, chamber pressure is 0.15Pa, sputtering current is 0.16A

[0062] S3: placing the titanium film slide in a 0.2 mol / L sodium hydroxide solution, activating at 50°C for 30 min, washing, purging with nitrogen, and drying to obtain a hot alkali activated slide;

[0063] S4: Place 1mmol sodium alginate in a reaction container, add pH 7.2 phosphate buffer solution, stir evenly, add 3mmol 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and 3mmol N-hydroxysuccinimide, stir to activate, add 2mmol mono-(6-amino-6-deoxy)-β-cyclodextrin solution, stir evenly, react at room temperature for 24h, purify, freeze-dry, and obtain cyclodextrin-modified sodium alginate; add sodium periodate solution to 1.2wt% cyclodextrin-modified sodium alginate solution, stir evenly in a dark environment, react for 24h, add ethylene glycol to terminate the reaction, purify, and obtain dialdehyde β-cyclodextrin-modified oxidized sodium alginate solution; wherein the molar ratio of sodium periodate in the sodium periodate solution: cyclodextrin-modified sodium alginate: ethylene glycol in the cyclodextrin-modified sodium alginate solution is 0.5:1:0.5;

[0064] S5: soaking the hot alkali activated glass slide in a 1 mg / mL poly-L-lysine solution, taking it out and washing it with deionized water to obtain a layer of poly-L-lysine film; dropping 1 mg / mL dialdehyde β-cyclodextrin modified oxidized sodium alginate solution on the poly-L-lysine film, spin coating, washing, and drying to obtain a prepared low-detachment rate glass slide; wherein the thickness of the dialdehyde β-cyclodextrin modified oxidized sodium alginate layer is 0.25 μm;

[0065] S6: Add 0.5 mg / mL poly-D-lysine solution onto the prepared low detachment rate glass slide, spin coat, wash, then add 1 mg / mL dialdehyde β-cyclodextrin modified oxidized sodium alginate solution, spin coat, wash, repeat the above poly-L-lysine solution and dialdehyde β-cyclodextrin modified oxidized sodium alginate solution for one spin coating, the coating thickness of one spin coating is 0.25 μm; repeat the spin coating steps 8 times to prepare a low detachment rate adhesive glass slide.

[0066] Comparative Example 2: A method for preparing an adhesive glass slide with a low detachment rate: S1: After the glass slide is washed with deionized water, it is placed in a potassium permanganate solution and soaked for 6 hours, taken out and washed with deionized water, and finally ultrasonically washed with deionized water, and dried at 80°C to obtain a pretreated glass slide;

[0067] S2: The pretreated glass slide was used as the substrate, and the pattern grating template was placed, and magnetron sputtering was performed to obtain a titanium thin film glass slide; wherein the magnetron sputtering target material was Ti target, the magnetron sputtering time was 7 min, and the vacuum degree was 5.5×10 -4 Pa, argon flow rate is 20sccm, chamber pressure is 0.15Pa, sputtering current is 0.16A

[0068] S3: placing the titanium film slide in a 0.2 mol / L sodium hydroxide solution, activating at 50°C for 30 min, washing, purging with nitrogen, and drying to obtain a hot alkali activated slide;

[0069] S4: 1 mmol of carboxymethyl chitosan is placed in a reaction container, a pH 7.2 phosphate buffer solution is added, the mixture is stirred evenly, 3 mmol of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and 3 mmol of N-hydroxysuccinimide are added, the mixture is stirred for activation, 2 mmol of mono-(6-amino-6-deoxy)-β-cyclodextrin solution is added, the mixture is stirred evenly, the reaction is carried out at room temperature for 12 h, the mixture is purified, and the mixture is freeze-dried to obtain cyclodextrin-modified chitosan;

[0070] S5: Soak the hot alkali activated glass slide in a 1 mg / mL poly-L-lysine solution, remove it and wash it with deionized water to obtain a layer of poly-L-lysine film; drip 1 mg / mL cyclodextrin modified chitosan solution on the poly-L-lysine film, spin coat, wash, and dry to obtain a prepared low-detachment rate glass slide; wherein the thickness of the dialdehyde β-cyclodextrin modified oxidized sodium alginate layer is 0.25 μm;

[0071] S6: Add 0.5 mg / mL poly-L-lysine solution on the prepared low detachment rate slide glass, spin coat, wash, then add 1 mg / mL cyclodextrin modified chitosan solution, spin coat, wash, repeat the above poly-L-lysine solution and cyclodextrin modified chitosan solution for one spin coating, the coating thickness of one spin coating is 0.25 μm; repeat the spin coating steps 8 times to prepare a low detachment rate adhesive slide glass.

[0072] Comparative Example 3: A method for preparing an adhesive glass slide with a low detachment rate: S1: After washing the glass slide with deionized water, soak it in a potassium permanganate solution for 6 hours, take it out and wash it with deionized water, and finally ultrasonically clean it with deionized water, and dry it at 80°C to obtain a pretreated glass slide;

[0073] S2: The pretreated glass slide was used as the substrate, and the pattern grating template was placed, and magnetron sputtering was performed to obtain a titanium thin film glass slide; wherein the magnetron sputtering target material was Ti target, the magnetron sputtering time was 7 min, and the vacuum degree was 5.5×10 -4 Pa, argon flow rate is 20sccm, chamber pressure is 0.15Pa, sputtering current is 0.16A

[0074] S3: placing the titanium film slide in a 0.2 mol / L sodium hydroxide solution, activating at 50°C for 30 min, washing, purging with nitrogen, and drying to obtain a hot alkali activated slide;

[0075] S4: adding sodium periodate solution to 1.0wt% sodium alginate solution, stirring evenly in a dark condition, reacting for 24h, adding ethylene glycol to terminate the reaction, purifying, and freeze-drying to obtain oxidized sodium alginate; wherein the molar ratio of sodium periodate in the sodium periodate solution: sodium alginate in the sodium alginate solution: ethylene glycol is 0.5:1:0.5; placing 1mmol of oxidized sodium alginate in a reaction container, adding pH5.72-morpholineethanesulfonic acid solution, stirring evenly, adding 3mmol1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and 3mmolN-hydroxysuccinimide, stirring for activation, adding 2mmol mono-(6-amino-6-deoxy)-β-cyclodextrin solution, stirring evenly, reacting at room temperature for 24h, purifying, and freeze-drying to obtain cyclodextrin-modified oxidized sodium alginate;

[0076] S5: soaking a hot alkali activated glass slide in a 1 mg / mL poly-L-lysine solution, taking it out and washing it with deionized water to obtain a layer of poly-L-lysine film; dropping a 1 mg / mL cyclodextrin-modified oxidized sodium alginate solution on the poly-L-lysine film, spin coating, washing, and drying to obtain a prepared low-detachment rate glass slide; wherein the thickness of the dialdehyde β-cyclodextrin-modified oxidized sodium alginate layer is 0.25 μm;

[0077] S6: Add 0.5 mg / mL poly-L-lysine solution on the prepared low detachment rate glass slide, spin coat, wash, then add 1 mg / mL cyclodextrin modified oxidized sodium alginate solution, spin coat, wash, repeat the above poly-L-lysine solution and cyclodextrin modified oxidized sodium alginate solution for one spin coating, the coating thickness of one spin coating is 0.25 μm; repeat the spin coating steps 8 times to prepare a low detachment rate adhesive glass slide.

[0078] Comparative Example 4: A method for preparing an adhesive glass slide with a low detachment rate: S1: After washing the glass slide with deionized water, soak it in a potassium permanganate solution for 6 hours, take it out and wash it with deionized water, and finally ultrasonically clean it with deionized water, and dry it at 80°C to obtain a pretreated glass slide;

[0079] S2: Place 1mmol sodium alginate in a reaction container, add pH 7.2 phosphate buffer solution, stir evenly, add 3mmol 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and 3mmol N-hydroxysuccinimide, stir to activate, add 2mmol mono-(6-amino-6-deoxy)-β-cyclodextrin solution, stir evenly, react at room temperature for 24h, purify, freeze-dry, and obtain cyclodextrin-modified sodium alginate; add sodium periodate solution to 1.2wt% cyclodextrin-modified sodium alginate solution, stir evenly in a dark environment, react for 24h, add ethylene glycol to terminate the reaction, purify, and obtain dialdehyde β-cyclodextrin-modified oxidized sodium alginate solution; wherein the molar ratio of sodium periodate in the sodium periodate solution: cyclodextrin-modified sodium alginate: ethylene glycol in the cyclodextrin-modified sodium alginate solution is 0.5:1:0.5;

[0080] S3: Soak the pretreated glass slide in a 1 mg / mL poly-L-lysine solution, remove and wash with deionized water to obtain a layer of poly-L-lysine film; drip 1 mg / mL dialdehyde β-cyclodextrin modified oxidized sodium alginate solution on the poly-L-lysine film, spin coat, wash, and dry to obtain a preparatory low-detachment rate glass slide; wherein the thickness of the dialdehyde β-cyclodextrin modified oxidized sodium alginate layer is 0.25 μm;

[0081] S4: drip 0.5 mg / mL poly-L-lysine solution onto the prepared low-detachment rate glass slide, spin-coat, wash, then drip 1 mg / mL dialdehyde β-cyclodextrin modified oxidized sodium alginate solution, spin-coat, wash, repeat the above poly-L-lysine solution and dialdehyde β-cyclodextrin modified oxidized sodium alginate solution for one spin coating, the coating thickness of one spin coating is 0.25 μm; repeat the spin coating steps 8 times to prepare a low-detachment rate adhesive glass slide

[0082] Experiment: Cell adhesion assay: Human skin fibroblasts seeded on low-detachment rate adhesion slides were incubated at 37°C, 5% CO 2 After culturing in an incubator for 6 hours, the upper cell culture medium was aspirated, and the sample was washed 3 times with PBS buffer solution to remove the non-adherent cells; under light-proof conditions, FDA dye was added, and after staining for 20 minutes, the FDA dye was aspirated, and PBS buffer solution was added for washing 3 times, with a single washing time of 5 minutes and an oscillation speed of 300 rpm; the average cell density on the surface of the low detachment rate adhered slide was statistically analyzed using software;

[0083] Substrate adhesion test: Two low-detachment rate adhesive glass slides of the same size are bonded face to face and placed in a 45°C oven to keep constant weight; after constant weight, the two ends of the bonded glass substrate are clamped on a tensile tensile tester and the test is carried out at a tensile speed of 50 m / min until the two glass slides are separated. The same example sample is tested 5 times;

[0084] Slide surface WCA measurement experiment: The CA of the samples of Example 1 and Comparative Example 5 was measured using an optical contact angle meter. Distilled water was dropped on 5 different positions of each sample and the average value was calculated.

[0085] Table 1 Cell adhesion / substrate adhesion experiment data table

[0086]

[0087]

[0088] Table 2 Contact angle experimental data

[0089] Contact angle / ° Example 1 25.9 Comparative Example 4 29.1

[0090] Conclusion: The low detachment rate adhesive slides prepared in Examples 1-5 have excellent hydrophilicity and cell adhesion properties; the poly-D-lysine used in Comparative Example 1 replaced the poly-L-lysine, resulting in a decrease in cell adhesion performance; the use of carboxymethyl chitosan instead of sodium alginate in Comparative Example 2 resulted in a decrease in cell adhesion performance; the swapping of the order of sodium periodate and grafted β-cyclodextrin in Comparative Example 3 resulted in the lack of a dialdehyde structure in β-cyclodextrin, resulting in a reduction in the Schiff base structure and a decrease in various properties; the lack of magnetron sputtering in Comparative Example 4 to prepare the titanium thin film layer resulted in a decrease in hydrophilicity, cell adhesion and substrate adhesion properties.

[0091] Finally, it should be noted that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for preparing an adhesive glass slide with low detachment rate, characterized in that: The method comprises the following steps: S1: soaking a hot alkali activated glass slide in a poly-L-lysine solution, taking out and washing with deionized water to obtain a layer of poly-L-lysine film; The dialdehyde β-cyclodextrin modified oxidized sodium alginate solution is dripped onto the poly-L-lysine film, spin-coated, washed, and dried to obtain a prepared low-detachment rate slide glass; S2: the poly-L-lysine solution is dripped onto the prepared low-detachment rate slide glass, spin-coated, washed, and then the dialdehyde β-cyclodextrin modified oxidized sodium alginate solution is dripped, spin-coated, washed, and the poly-L-lysine solution and the dialdehyde β-cyclodextrin modified oxidized sodium alginate solution are repeated as one spin coating, and the spin coating steps are repeated n times to prepare a low-detachment rate adhesive slide glass; The preparation method of the hot alkali activated glass slide comprises the following steps: step (1): washing the glass slide with deionized water, placing it in a potassium permanganate solution for 4-8 hours, taking it out and washing it with deionized water, finally ultrasonically washing it with deionized water, and drying it at 80-85° C. to obtain a pretreated glass slide; step (2): using the pretreated glass slide as a substrate, placing a pattern grating template, and magnetron sputtering to obtain a titanium film glass slide; step (3): placing the titanium film glass slide in a sodium hydroxide solution, activating it, washing it, purging it with nitrogen, and drying it to obtain a hot alkali activated glass slide; The preparation method of the dialdehyde β-cyclodextrin modified oxidized sodium alginate solution comprises the following steps: placing sodium alginate in a reaction container, adding a buffer solution and stirring evenly, adding a cross-linking auxiliary agent and stirring to activate, adding a mono-(6-amino-6-deoxy)-β-cyclodextrin solution and stirring evenly, reacting at room temperature for 22-26 hours, purifying, and freeze-drying to obtain cyclodextrin modified sodium alginate; adding a sodium periodate solution to the cyclodextrin modified sodium alginate solution, stirring evenly in a light-proof condition, reacting for 20-24 hours, adding ethylene glycol to terminate the reaction, and purifying to obtain a dialdehyde β-cyclodextrin modified oxidized sodium alginate solution.

2. The method for preparing a low detachment rate adhesive glass slide according to claim 1, characterized in that: In the step (2), the magnetron sputtering target is a Ti target, the magnetron sputtering time is 7-9 min, and the vacuum degree is 5.5×10 -4 -6.0×10 -4 Pa, argon flow rate is 20-30sccm, chamber pressure is 0.15-0.18Pa, and sputtering current is 0.16-0.19A.

3. The method for preparing a low detachment rate adhesive glass slide according to claim 1, characterized in that: In the step (3), the concentration of the sodium hydroxide solution is 0.1-0.2 mol / L; the activation temperature is 45-50° C., and the activation time is 25-30 min.

4. The method for preparing a low detachment rate adhesive glass slide according to claim 1, characterized in that: The buffer solution is a phosphate buffer solution with a pH of 7.2-7.3; the cross-linking auxiliary agent is a mixture of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide in a molar ratio of 1:1; the solvent in the cross-linking auxiliary agent and mono-(6-amino-6-deoxy)-β-cyclodextrin solution is a buffer solution; the molar ratio of the sodium alginate: the cross-linking auxiliary agent is (1-1.2): (6-6.4); the molar ratio of the sodium alginate: the mono-(6-amino-6-deoxy)-β-cyclodextrin is (1-1.2): (2-2.4).

5. The method for preparing a low detachment rate adhesive glass slide according to claim 1, characterized in that: The concentration of the cyclodextrin modified sodium alginate solution is 1.2-1.5 wt %.

6. The method for preparing a low detachment rate adhesive glass slide according to claim 1, characterized in that: In the step S1, the concentration of the poly-L-lysine solution is 1-2 mg / mL, and the concentration of the dialdehyde β-cyclodextrin modified oxidized sodium alginate solution is 1-2 mg / mL.

7. The method for preparing a low detachment rate adhesive glass slide according to claim 1, characterized in that: In the step S2, the concentration of the poly-L-lysine solution is 0.5-1 mg / mL, and the concentration of the dialdehyde β-cyclodextrin modified oxidized sodium alginate solution is 1-2 mg / mL; the spin coating is repeated n times n=8-12 times, and the coating thickness of one spin coating is 0.25-0.3 μm.

8. An adhesive glass slide with low detachment rate prepared by the method for preparing an adhesive glass slide with low detachment rate according to any one of claims 1 to 7.

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