Forming structure and forming method of film

Abstract

The invention provides a film forming structure and method, the film forming structure comprises a support body, the support body is filled with a first buffer solution, the surface of the first buffer solution is provided with a film solution, the surface of the film solution is provided with a second buffer solution, and the second buffer solution is provided with a first buffer solution and a second buffer solution. The osmotic pressure of the first buffer solution is greater than that of the second buffer solution. According to the forming structure and the forming method, the support of a specific structure can be quickly responded, operable windows of the support are increased, controllability and uniformity are achieved, the film forming period is shortened, and corresponding cost is saved.

Description

Film forming structure and forming method Technical Field

[0001] The present application belongs to the field of membrane technology and relates to a membrane forming structure and a forming method. Background Art

[0002] At present, in the preparation of bilayer membranes on the market, the technical points focus on the method of brushing the membrane, that is, considering optimizing the support or its array structure, surface properties or adjusting the ratios of the membrane reagents to achieve brushing the membrane on the support. Correspondingly, in the process of forming the bilayer, excluding the reasons of its own performance, it will also be affected by other external factors. However, in promoting the formation of bilayer membranes, there is a lack of operating methods, and most of them rely entirely on its spontaneous process, resulting in a long film formation cycle and poor uniformity, which will bring large batch-to-batch differences.

[0003] The main disadvantages of the existing bilayer membrane forming process are: 1. Poor controllability and excessive reliance on its spontaneity, which is significantly affected by external factors. For example, for a support with a certain structure (small diameter), the membrane forming cycle will be lengthened and lack controllability; 2. Poor uniformity. First of all, there are differences between objects, and the subsequent operation procedures will greatly bring differences, which ultimately leads to obvious differences between individual results of the product, making it inconvenient for subsequent operation and processing; 3. It increases the difficulty of production in disguise and increases its cost.

[0004] Summary of the Invention

[0005] The present application provides a film forming structure and forming method, which can also quickly respond to supports of specific structures, increase their operable windows, achieve controllability and uniformity, shorten the film forming cycle, and save corresponding costs.

[0006] One of the purposes of the present application is to provide a membrane forming structure, which includes a support body, the interior of the support body is filled with a first buffer solution, a membrane solution is provided on the surface of the first buffer solution, and a second buffer solution is provided on the surface of the membrane solution (more specifically, the other side surface), and the osmotic pressure of the first buffer solution is greater than the osmotic pressure of the second buffer solution.

[0007] In this application, the process of forming a bilayer membrane for amphiphilic molecules is a self-assembly process driven by the hydrophobic effect, which is determined by the properties of the material itself, and the degree of spontaneity of different membrane materials is different. However, for the same membrane material, the self-assembly process will be affected by its surrounding environmental factors, such as the viscosity, ratio, temperature and humidity environment of the solvent. The film-forming structure described in this application utilizes the barrier principle of the water permeability of the membrane material to macromolecules, so that the osmotic pressure value below the membrane is greater than that above, resulting in a deficit on both sides. Water molecules will move to the part with high osmotic pressure, causing the membrane to have an upward convex pulling force. It is conducive to the outward movement of the solvent in the membrane solution, accelerating the process of forming a bilayer membrane.

[0008] As a preferred technical solution of the present application, the osmotic pressure of the first buffer solution is 180-400 mOsmol / kg, such as 374 mOsmol / kg, 281 mOsmol / kg or 187 mOsmol / kg.

[0009] As a preferred technical solution of the present application, the first buffer solution includes a salt solution of potassium chloride, sodium chloride, potassium ferrocyanide, potassium ferrocyanide, sodium ferrocyanide, sodium ferrocyanide, sodium carbonate, or sodium bicarbonate. In some more preferred embodiments, depending on the actual requirements of subsequent steps, the first buffer solution may optionally contain other components, such as hepes, hepes-Na-hepes, or K2HPO4-KH2PO4.

[0010] As a preferred technical solution of the present application, the osmotic pressure of the second buffer solution is 140 to 300 mOsmol / kg, such as 289 mOsmol / kg, 154 mOsmol / kg or 145 mOsmol / kg.

[0011] As a preferred technical solution of the present application, the second buffer solution includes a salt solution of potassium chloride, sodium chloride, potassium ferrocyanide, potassium ferrocyanide, sodium ferrocyanide, sodium ferrocyanide, sodium carbonate, and sodium bicarbonate. In some more preferred embodiments, according to the actual needs of the subsequent steps, the second buffer solution may optionally contain other components, such as hepes, hepes-Na-hepes, or K2HPO4-KH2PO4. In the present application, the difference in osmotic pressure between the first buffer solution and the second buffer solution can be used to control the film forming process. When the osmotic pressure difference is increased at a specific location, the formation of the bilayer membrane can be accelerated to obtain a membrane with a larger capacitance value, but too large a capacitance value will lead to membrane rupture. When the osmotic pressure difference is reduced, the effect of accelerating the formation of the bilayer membrane is not very obvious, and a membrane with a slightly larger capacitance value is obtained, but the membrane is more stable.

[0012] As a preferred technical solution of the present application, the membrane solution is prepared into a bilayer membrane by self-assembly.

[0013] As a preferred technical solution of the present application, the membrane solution includes an amphiphilic membrane material and a solvent.

[0014] The amphiphilic membrane material includes any one of DPHPC, DPHPG, DPPC, POPC or DOPE, or a combination of at least two thereof.

[0015] Among them, DPHPC refers to diphytanoylphosphatidylcholine 1,2-diphytanoyl-sn-glycero-3-phosphocholine;

[0016] DPHPG refers to 1,2-diphytanoyl-sn-glycero-3-phospho-(1'-rac-glycerol);

[0017] DPPC refers to 1,2-dipalmitoyl-sn-glycero-3-phosphocholine;

[0018] POPC refers to phosphatidylcholine (1-palmitoyl-2-oleoyl-glycero-3-phosphocholine);

[0019] DOPE refers to 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine.

[0020] The solvent includes one or more of decane, dodecane, and n-octane and any one of AR20, AR30, and PDMS-9 or a combination of at least two thereof.

[0021] Among them, AR20 refers to silicone oil with a viscosity of 20mPa.s (25°C), also known as polyphenyl-methylsiloxane.

[0022] In the present application, the types and substances of the first buffer solution, the second buffer solution and the membrane solution are merely possible examples, and can be specifically selected according to the preparation requirements of the membrane, and no further limitation is given.

[0023] A second object of the present application is to provide a film forming method, which includes forming the above-mentioned forming structure.

[0024] As a preferred technical solution of this application, the molding method includes:

[0025] Under vacuum conditions, a first buffer solution is injected into the support, and a second buffer solution is used to push the membrane solution along the support to cover the surface of the first buffer solution, and the solution is allowed to stand to obtain a bilayer membrane.

[0026] As a preferred technical solution of this application, the molding method includes:

[0027] A first buffer solution is injected into the support body, and the membrane solution is pushed along the support body to cover the surface of the first buffer solution using the first buffer solution. After the membrane solution is self-assembled and in a stable state, a second buffer solution is used to replace the first buffer solution above the membrane solution, and the solution is allowed to stand to obtain a bilayer membrane.

[0028] In the present application, the stable state after the membrane solution is self-assembled and in a stable state refers to a state in which the membrane solution on the support has formed a bilayer, which is manifested by the capacitance value reaching the theoretical value, which is greater than 10pF and less than 200pF.

[0029] In this application, the process parameter window of the film brushing stage is relatively narrow. Under the condition that the film brushing process parameters are not changed, the second forming method can be adopted. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] FIG1 is a schematic diagram of a membrane optimization scheme for narrow process parameters according to the method for optimizing the osmotic pressure principle of the present application;

[0031] FIG2 is a schematic diagram of a film forming optimization scheme for ductile membrane materials using the osmotic pressure principle optimization method of the present application;

[0032] FIG3 is a schematic diagram illustrating the principle of this application;

[0033] FIG4 is a confocal laser scanning image of an unqualified film formed under isotonic pressure;

[0034] FIG5 is a confocal laser scanning image of an unqualified film formed under isotonic pressure after being processed by an optimization method;

[0035] Figure 6 is a membrane capacitance map after the membrane was formed and left to stand for 24 hours under isotonic pressure;

[0036] FIG7 is a distribution diagram of membrane capacitance values ​​after the membrane is formed and left to stand for 24 hours under osmotic pressure;

[0037] Figure 8 is a diagram of the membrane capacitance map after the membrane was formed and left to stand for 24 hours under isotonic pressure and then processed by the optimization method;

[0038] FIG9 is a distribution diagram of the membrane capacitance value after the membrane is formed and left to stand for 24 hours under isotonic pressure and treated by the optimization method;

[0039] FIG10 is a graph showing the change in membrane capacitance over time after the membrane was formed and left to stand for 24 hours under isotonic pressure and before and after treatment using the optimized method;

[0040] FIG11 is a graph showing the change in membrane capacitance over time during the static film formation process under non-isotonic pressure in Example 3;

[0041] FIG12 is a graph showing the change in membrane capacitance over time in Example 4 after forming the membrane under isotonic pressure and standing for 24 hours before and after treatment by the optimized method;

[0042] FIG13 is a graph showing the change in membrane capacitance over time of Example 5 after the membrane is formed at isotonic pressure and allowed to stand for 24 hours before and after treatment by the optimized method.

[0043] The following is a further detailed description of the present application. However, the following examples are merely simplified examples of the present application and do not represent or limit the scope of protection of the present application. The scope of protection of the present application shall be subject to the claims. DETAILED DESCRIPTION

[0044] The technical solution of the present application will be further explained below with reference to the accompanying drawings and through specific implementation methods.

[0045] To better illustrate the present application and facilitate understanding of the technical solution of the present application, typical but non-limiting embodiments of the present application are as follows:

[0046] The reagents used in the specific implementation of this application are as follows:

[0047] First buffer solution: 500 mL; 200 mMol / L KCl, 1 mMol / L HEPES, 1 mMol / L EDTA, osmotic pressure 374 mOsmol / kg, designated as buffer 1 in the following examples. To 100 mL of buffer 1, 0.02% by weight of rhodamine dye was added to obtain a solution designated as buffer 1-1 in the following examples.

[0048] The second buffer solution: 500 mL; 155 mMol / L KCL, 1 mMol / L HEPES, 1 mMol / L EDTA, osmotic pressure of 289 mOsmol / kg, named buffer 2 in the following examples.

[0049] Membrane solution: 3 mL of 10 mg / mL DPHPC, designated as Reagent A in the following examples. 1 mL of Reagent A was added with 0.01% by weight of fluoroborane pyrrole dye to obtain a solution designated as Reagent A-1 in the following examples.

[0050] The support structure is shown in Figure 1.

[0051] Example 1

[0052] This embodiment provides a film forming method, the forming method comprising:

[0053] Buffer 1-1 was injected into the support, and buffer 2 was used to push the membrane solution (reagent A-1) along the support to cover the buffer 1-1. After standing for 24 hours, the membrane solution was self-assembled and in a stable state to obtain a bilayer membrane.

[0054] Example 2

[0055] This embodiment provides a film forming method, the forming method comprising:

[0056] Buffer 1 was injected into the support, and the membrane solution (reagent A) was pushed along the support using buffer 1 to cover the buffer 1. After standing for 24 hours, the membrane solution was self-assembled and in a stable state, and then buffer 2 was used to replace the buffer 1 above the membrane solution. The membrane was allowed to stand for 30 minutes to obtain a bilayer membrane.

[0057] Comparative Example 1

[0058] This comparative example provides a film forming method, the forming method comprising:

[0059] Buffer 1-1 was injected into the support, and the membrane solution (reagent A-1) was pushed along the support using buffer 1 to cover the buffer 1-1. After the membrane solution was self-assembled and in a stable state, it was allowed to stand for 24 hours.

[0060] Comparative Example 2

[0061] This comparative example provides a film forming method, the forming method comprising:

[0062] Buffer 1 was injected into the support, and the membrane solution (reagent A) was pushed along the support using buffer 1 to cover the buffer 1. After the membrane solution was self-assembled and in a stable state, it was allowed to stand for 24 hours.

[0063] The film prepared in Comparative Example 1 was placed under a confocal laser scanning microscope, using a 20x10 lens combination and a scanning speed of 5μm / step to scan the film on the support. Figure 3 shows a thicker film layer above buffer 1-1 in the support cavity. Buffer 2 was used to replace the upper buffer 1 layer in the film-forming structure of Comparative Example 1 (i.e., the bilayer film prepared in Example 1), and a scan was performed under the scanning microscope to obtain Figure 4. The image shows that the oily solvent in the film solution has been repelled to the outside, and a bilayer has been formed.

[0064] The bilayer membrane obtained in Comparative Example 2 was placed under an instrument for testing membrane capacitance, and its membrane capacitance value was detected. Its map is shown in Figure 6 below, and the capacitance distribution diagram is shown in Figure 7 below. It was found that after the membrane was left to stand for 24 hours under isotonic pressure, its membrane capacitance was uneven and there were two peaks with large deviations, at 5pF and 24pF respectively, and 225 channels did not form a bilayer. Buffer 1 was replaced with buffer 2, and the bilayer membrane prepared in Example 2 was left to stand for 30 minutes to observe its capacitance change over time. After replacement for 1 hour, its map was shown in Figure 8 below, and the capacitance distribution diagram is shown in Figure 9 below. It was found that except for the 3 channels that did not form the upper membrane, only 1 channel among the 225 channels that did not form a bilayer had not yet formed. Therefore, 1024 channels were successfully improved from 796 to 1020. The qualified rate was improved from 77.73% to 99.61%, and the uniformity was good. The capacitance change diagram over time is shown in Figure 10.

[0065] Example 3

[0066] This embodiment provides a film forming method, the forming method comprising:

[0067] Under vacuum conditions, buffer 1 was injected into the support. Buffer 2 was used to push the membrane solution (reagent A) along the support to cover the surface of buffer 1. The capacitance was observed over time. Under non-isotonic conditions, the osmotic pressure difference accelerated the self-assembly process. Results showed that after 4 hours of standing, a large number of channels began to self-assemble into a bilayer membrane. After 12 hours, 90% of the 1024 channels had completed their self-assembly into a bilayer membrane. A graph of the capacitance over time is shown in Figure 11.

[0068] Example 4

[0069] This example provides a membrane formation method. The conditions are identical to those in Example 2, except that the osmotic pressure of buffer 1 is 281 mOsmol / kg (designated buffer 1' in this example) and the osmotic pressure of buffer 2 is 154 mOsmol / kg (designated buffer 2' in this example). Observation of the change in capacitance over time revealed that, under isotonic conditions, approximately 200 (20%) channels failed to form a bilayer membrane. The osmotic pressure difference between the upper replacement buffer 1' and the lower replacement buffer 2' was significantly greater than that in Example 2. After 30 minutes of replacement, 99% of the 1024 channels had completed self-assembly to form a bilayer membrane. A graph of the change in capacitance over time is shown in Figure 12.

[0070] Example 5

[0071] This example provides a membrane forming method. The forming conditions are identical to those of Example 2, except that the osmotic pressure of buffer 1 is 187 mOsmol / kg (designated buffer 1″ in this example) and the osmotic pressure of buffer 2 is 145 mOsmol / kg (designated buffer 2″ in this example). Observation of the change in capacitance over time revealed that, under isotonic conditions, approximately 200 (20%) channels failed to form a bilayer membrane. The osmotic pressure difference between the upper replacement buffer 1″ and the lower buffer 2″ was smaller than that in Example 2. After 30 minutes of replacement, 99% of the 1024 channels had completed their self-assembly process, resulting in a bilayer membrane. A graph of the change in capacitance over time is shown in Figure 13.

[0072] In summary, this application has the following beneficial effects:

[0073] (1) The present application provides a membrane forming structure and forming method, which utilizes the principle of osmotic pressure to accelerate the process of amphiphilic materials forming a bilayer membrane, while also greatly shortening the production cycle, saving costs, and improving production efficiency;

[0074] (2) The present application provides a membrane forming structure and forming method, which can achieve controllability of the membrane forming process by controlling the osmotic pressure difference;

[0075] (3) The present application provides a film forming structure and forming method, which have a wide range of application scenarios and a large product window;

[0076] (4) The present application provides a film forming structure and forming method, wherein the film forming structure and forming method have good uniformity and a high qualified rate per unit time.

[0077] The applicant declares that while the above-mentioned embodiments are used to illustrate the detailed structural features of the present application, the present application is not limited to the above-mentioned detailed structural features, which does not mean that the present application must rely on the above-mentioned detailed structural features in order to be implemented. Those skilled in the art should understand that any improvements to the present application, equivalent replacements for selected components, additions of auxiliary components, and selection of specific methods, etc., fall within the scope of protection and disclosure of the present application.

[0078] The preferred embodiments of the present application are described in detail above. However, the present application is not limited to the specific details of the above embodiments. Within the technical concept of the present application, various simple modifications can be made to the technical solution of the present application, and these simple modifications all fall within the scope of protection of the present application.

[0079] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any appropriate manner unless there is any contradiction. In order to avoid unnecessary repetition, this application will not further describe various possible combinations.

[0080] In addition, the various implementation methods of the present application may be arbitrarily combined, and as long as they do not violate the concept of the present application, they should also be regarded as the contents disclosed in the present application.

Claims

1. A membrane forming structure, comprising a support body, wherein the support body is filled with a first buffer solution, a membrane solution is disposed on the surface of the first buffer solution, and a second buffer solution is disposed on the surface of the membrane solution, wherein the osmotic pressure of the first buffer solution is greater than the osmotic pressure of the second buffer solution.

2. The molding structure according to claim 1, wherein: The osmotic pressure of the first buffer solution is 180-400 mOsmol / kg.

3. The forming structure according to claim 1 or 2, wherein: The first buffer solution includes a salt solution of potassium chloride, sodium chloride, potassium ferrocyanide, potassium ferrocyanide, sodium ferrocyanide, sodium ferrocyanide, sodium carbonate or sodium bicarbonate.

4. The molded structure according to claim 3, wherein the first buffer solution further comprises hepes, hepes-Na-hepes, or K2HPO4-KH2PO4.

5. The molding structure according to any one of claims 1 to 4, wherein: The osmotic pressure of the second buffer solution is 140 to 300 mOsmol / kg.

6. The molding structure according to any one of claims 1 to 5, wherein: The second buffer solution includes a salt solution of potassium chloride, sodium chloride, potassium ferrocyanide, potassium ferrocyanide, sodium ferrocyanide, sodium ferrocyanide, sodium carbonate or sodium bicarbonate.

7. The molded structure according to claim 6, wherein the second buffer solution further comprises hepes, hepes-Na-hepes, or K2HPO4-KH2PO4.

8. The molding structure according to any one of claims 1 to 7, wherein: The membrane solution is used to prepare a bilayer membrane through self-assembly.

9. The molding structure according to any one of claims 1 to 8, wherein: The membrane solution includes an amphiphilic membrane material; The amphiphilic membrane material includes any one of DPHPC, DPHPG, DPPC, POPC or DOPE, or a combination of at least two thereof.

10. The molding structure according to claim 9, wherein: The membrane solution also includes a solvent.

11. The molding structure according to claim 10, wherein: The solvent includes one or more of Decane, dodecane, and n-octane and any one of AR20, AR30, and PDMS-9 or a combination of at least two thereof.

12. A method for forming a film, comprising forming the formed structure according to any one of claims 1 to 11.

13. The molding method according to claim 12, wherein: The molding method comprises: Under vacuum conditions, a first buffer solution is injected into the support, and a second buffer solution is used to push the membrane solution along the support to cover the surface of the first buffer solution, and the solution is allowed to stand to obtain a bilayer membrane.

14. The molding method according to claim 12, wherein: The molding method comprises: A first buffer solution is injected into the support body, and the membrane solution is pushed along the support body to cover the surface of the first buffer solution using the first buffer solution. After the membrane solution is self-assembled and in a stable state, a second buffer solution is used to replace the first buffer solution above the membrane solution, and the solution is allowed to stand to obtain a bilayer membrane.

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