CN102072971A - Method for preparing lipid bilayer membrane in liquid phase in situ - Google Patents
Method for preparing lipid bilayer membrane in liquid phase in situ Download PDFInfo
- Publication number
- CN102072971A CN102072971A CN2009101992319A CN200910199231A CN102072971A CN 102072971 A CN102072971 A CN 102072971A CN 2009101992319 A CN2009101992319 A CN 2009101992319A CN 200910199231 A CN200910199231 A CN 200910199231A CN 102072971 A CN102072971 A CN 102072971A
- Authority
- CN
- China
- Prior art keywords
- vesica
- liquid
- solution
- liquid pool
- injector
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Sampling And Sample Adjustment (AREA)
Abstract
The invention belongs to the technical field of preparation of membranes, and relates to a method for preparing a lipid bilayer membrane in a liquid phase in situ. The formed bilayer membrane can be observed and researched by an atomic force microscope. In the method, the preparation method in the prior art is improved, so a formation process and a surface topography of the membrane are monitored by the atomic force microscope in real time; the liquid-phase environment of the membrane can be changed in situ through a micro-injector and a circulating pump; chemical and physical adsorption, a chemical reaction, crystal growth and the like on the membrane surface are controlled; and the method can also be used for preparing and modifying other two amphiphilic molecular membranes and controlling the characteristics of a medium of the membrane. The method provides a feasible and reliable approach for researching a biomembrane surface under a physiological environment and various physical and chemical processes of a material surface under a liquid-phase environment in real time in situ from a molecular-resolution size, and has extremely important research and application value.
Description
Technical field
The invention belongs to the membrane preparation technology field, relate to the preparation method of the lipid bilayer membrane of solid surface support, be specifically related to a kind of method at liquid phase made acid-stable in situ lipid bilayer membrane.
Background technology
(Supported Lipid Bilayer SLB), is one of biomembranous principal mode of model to the plane lipid bilayer membrane that solid surface supports.By the proportion of composing of control lipid bilayer and the influence factors such as medium character around the film, people can study biological membrane to the response of outside environmental change and the inherent mechanism of particular physiological process quantitatively by qualitative the reaching of atomic force microscope (AFM).The method for preparing the plane supporting film commonly used has spin-coating method, LB film transfer method and vesica fusion method three major types.Wherein, spin-coating method is to obtain multilayer lipid dry film by the spin coating instrument in the organic solution of the direct spin coating lipid of mica surface; LB film transfer method is by LB film instrument, the unimolecular layer membrane in the liquid bath is plated to solid surface such as mica and obtains individual layer lipid dry film.These two kinds of methods are convenient and swift, and are biological mobile but the adipose membrane that makes does not possess, and also can't obtain having the mobile molecule tunic of two dimension in the solution even be soaked in for a long time, thereby be of limited application.Described vesica fusion method is a kind of method for preparing lipid bilayer membrane that proposes recently, utilize the spontaneous characteristics of adjusting in the spontaneous fusion of solid surface such as mica of lipid vesica energy, impel lipid to form regular individual layer bilayer adipose membrane at solid surface, whole process of preparation is to occur in the liquid phase environment simultaneously, thereby the two dimension that can guarantee adipose membrane is mobile, can promote the research to replica.But at present all there is the following shortcoming in the various vesica fusion methods that propose: required chronic of (1) vesica spontaneous fusion film forming on solid surface, about 4～12 hours; (2) it is not high enough to obtain the probability of the better bimolecular tunic of effect; (3) in the process of the surperficial unnecessary vesica of flushing, usually can cause the damage of film; (4) in specific implementation process, when especially observing on transferring to the AFM sample stage, the film surface is often answered ingress of air and is lost flowability.In addition, all there are following two common problems in present various preparation methods: be difficult to the temperature that keeps required in operation and transfer process, especially for multicomponent system, may cause phase-splitting and the phase transformation vestige that can not eliminate; Any further modification or modification can not be realized, thereby in-situ modified and corresponding dynamics research can not be used for the film surface.
Therefore, study the method that the uniform and stable solid surface of a kind of convenient preparation supports lipid bilayer membrane, and realize monitoring in the solution environmental temperature and molion kind etc. in real time to the influence and the effect of membrane structure with AFM, be one can bigger promotion biological membrane research and have a problem of actual application value.
Prior art related to the present invention has:
1. Yang Fuyu edits. biological membrane [M]. and Beijing: scientific and technological publishing house, 2005.
2.W.Binder，V.Barragan，F.Menger.Domains?and?Rafts?in?Lipid?Membranes[J].Angew.Chem.Int.Ed.2003(42)：5802-5827.
3.S.Connell，D.Smith.The?atomic?force?microscope?as?a?tool?for?studying?phaseseparation?in?lipid?membranes.Molecular?Membrane?Biology，2006(23)：17-28.
4.E.Sackmann.Supported?membranes：Scientific?and?practical?applications[J].Science?1996(271)：43-48.
Summary of the invention
The objective of the invention is to propose the new preparation process of the lipid bilayer membrane that a kind of solid surface supports, be specifically related to a kind of method at liquid phase made acid-stable in situ lipid bilayer membrane.
The present invention quickens vesica by hot conditions and merges by in-situ preparing lipid bilayer membrane on the controllable temperature sample stage of atomic force microscope (AFM), realizes quick sample preparation.This method can also effectively be controlled the factor that the character of the residing temperature of duplicature, surrounding liquid medium etc. influences the film pattern simultaneously flexibly.
For achieving the above object, the invention provides a kind of method at liquid phase made acid-stable in situ lipid bilayer membrane, it is characterized in that, adopt in-situ high temperature vesica fusion method, in conjunction with atomic force microscope liquid pool punch block design sampling device, under hot conditions, merge vesica, at liquid phase made acid-stable in situ lipid bilayer membrane.
Among the present invention, on Controllable Temperature, the AFM sample stage, in the liquid pool, original position generates the support membrane of solid surface, and can cause the various physical-chemical reactions on film surface by the different liquid solution of displacement, monitor the variation of film surface topography in these processes in real time with atomic force microscope.
Among the present invention, on the sample stage of the controlled cold and hot scanner of atomic force microscope, setting up with the mica is the sample liquid pool device of substrate, inserts miniature thermometer and automated fluid injector.In the sample liquid pool, flow velocity, the consumption of the kind of solution and flushing displacement liquid are accurately controlled, with the character of assurance liquid phase and the reliability of concentration, do not damage the structure of film simultaneously.
In order to obtain stable smooth solid support film, present vesica fusion method is with a large amount of vesica solution (1-10 milliliter) that solid surface is fully excessive wetting earlier at normal temperatures mostly, and the fusion of carrying out not waiting in 4-12 hour obtains more smooth support membrane.
In view of the lipid vesica is formed by one or more lipids, every kind of lipid all has its specific phase transition temperature; In the time of on being in this temperature, lipid is in soft relatively liquid phase.Among the present invention, by the vesica solution of heating, make all lipids be in the temperature environment that is higher than transformation temperature, make the lipid vesica soft relatively, rigidity is less.The vesica of described softness is easy to merge after touching solid surface.Under the hot environment, vesica in the solution and solid surface collision probability increase simultaneously.These two kinds of effects will inevitably accelerating solid the formation of surface fat bimolecular support membrane.
Sampling device described in the present invention (shown in Figure 2 as Fig. 1) comprises, controllable temperature sample stage 0, liquid pool punch block 1, liquid pool inner chamber 2, liquid injector 3, thermometer 4, flexible pipe 5 and injector passage 6-8.
This device places on the controllable temperature sample stage 0 of atomic force microscope; wherein liquid pool punch block 1 bottom of commercial atomic force microscope is used to install the probe of atomic force microscope; there are three connectors the side; make things convenient for external solution to enter liquid pool inner chamber 2; or the connection external devices is surveyed the temperature in the liquid pool inner chamber 2; in addition, the protection of 0 type circle is arranged around the liquid pool inner chamber 2, can reduce moisture evaporation and mobile.
Sampling device of the present invention, wherein, the right side connector of liquid pool punch block 1 connects liquid injector 3, and middle connector connects thermometer 4, and the left side connector connects conventional hoses 5 as waste fluid channel.The liquid injector has three injector passage 6-8 to connect different types of liquid or solution, liquid or solution commonly used have Milli-Q water, various buffer solution, biological micromolecule aqueous solution and macromolecular aqueous solution etc., and can control selection, sample introduction speed and the sample size of solution by the valve of injector.Sampling device, thermometer and flexible pipe all can freely load and unload.
Particularly, the invention provides a kind of method, it is characterized in that, adopt said apparatus under hot conditions, to prepare the bilayer adipose membrane, comprise following concrete steps for atomic force microscope observation at liquid phase made acid-stable in situ lipid bilayer membrane:
(1) original position is injected vesica solution: before connecting sampling device, to there be solids such as the mica of smooth surface or cover glass to place on the controllable temperature sample stage of atomic force microscope, the liquid pool punch block is placed on the solid surface, in the liquid pool inner chamber, inject the lipid vesica solution of 60-100 μ l; Wherein the lipid concentration of lipid vesica solution is 0.5-2.0mg/ml, and the vesica size is less than 300nm.
(2) be installed into sampling device: sampling device is connected with the liquid pool punch block, closes the injector main valve, connect among three passage 6-8 of injector the syringe that 20-100ml tests required solution is housed; Install thermometer 4 and flexible pipe 5 simultaneously.
(3) high temperature merges: open sample stage temperature control power supply, and design temperature, wherein temperature range is 30-60 ℃, actual temp is set and is depended on used lipid in the system.When reaching design temperature with thermometer 4 readings, pick up counting, continue heating vesica solution 30-60 minute, on mica or cover glass surface, vesica merges the formation bimolecular film each other.
(4) original position is removed unnecessary vesica, obtains smooth bimolecular film.Close sample stage temperature control power supply, be cooled to room temperature.Open injector main valve and solution channel, the solution of 10-20ml is taken away the vesica that inner chamber does not merge by the liquid pool inner chamber, and waste liquid is derived the liquid pool inner chamber by flexible pipe.Mica or cover glass surface form smooth bimolecular tunic.
Advantage of the present invention is: adopt preparation method disclosed by the invention, compare with fusion method commonly used, time of fusion is short, is lower than 2 hours; Merging and removing in the process of unnecessary vesica and thoroughly got rid of air, guaranteed the planarization and the flowability of film; And this method has stronger dirigibility and application prospect, after forming the bimolecular tunic, also can pass through the solution kind and the solute quality of other channel change liquid pool inner chamber of injector, further observes the influence of various molecules to lipid bilayer membrane.
This method also can be used for preparation and other amphipathic molecule film of modification, as polymeric membrane etc., and equally can controlling diaphragm medium character of living in.
This method has been improved preparation solid surface support membrane spin-coating method, vesica fusion method and LB film transfer method commonly used, not only make the forming process of film and the surface topography of film be under the real-time monitoring of atomic force microscope, also can be by microsyringe and ebullator, original position changes the residing liquid phase environment of film, thereby realizes chemistry and physisorption, the chemical reaction on film surface and the crystal growth on film surface or the like on controlling diaphragm surface.The inventive method has very important research and using value for the various physical and chemical processes on research material surface under the liquid phase environment provide a feasible reliable approach.
The invention will be further described below in conjunction with drawings and Examples.
Description of drawings
The device side view of Fig. 1 for using among the present invention, wherein, 0 is the controllable temperature sample stage; 1 is the liquid pool punch block; 2 is the liquid pool inner chamber; 3 is the liquid injector.
The device vertical view of Fig. 2 for using among the present invention, wherein, 1 is the liquid pool punch block; 2 is the liquid pool inner chamber; 3 is the liquid injector; 4 is thermometer, measures temperature in 2; 5 is flexible pipe; 6-8: be the injector passage, can connect different liquids.
The AFM shape appearance figure of three component lipid (DOPC/ESM/Chol) films that Fig. 3 makes for embodiment 1, wherein appearance is of a size of 8 μ m * 8 μ m; The altitude scale of figure is 20nm.
The AFM shape appearance figure of three component lipid (DOPC/ESM/Chol) films that Fig. 4 makes for embodiment 2, wherein appearance is of a size of 8 μ m * 8 μ m; The altitude scale of figure is 10nm.
The AFM shape appearance figure of three component lipid (DOPC/ESM/Chol) the bimolecular tunics that Fig. 5 makes for embodiment 3, wherein appearance is of a size of 8 μ m * 8 μ m; The altitude scale of figure is 10nm.
The AFM shape appearance figure of three component lipid (DOPC/ESM/Chol) the bimolecular tunics that Fig. 6 makes for embodiment 4, wherein appearance is of a size of 10 μ m * 10 μ m; The altitude scale of figure is 10nm.
The AFM shape appearance figure of two component lipid (POPC/BSM/) the bimolecular tunics that Fig. 7 makes for embodiment 5, wherein appearance is of a size of 10 μ m * 10 μ m; The altitude scale of figure is 10nm.
The AFM shape appearance figure of two component lipid (DOPC/GM1) the bimolecular tunics that Fig. 8 makes for embodiment 6, wherein appearance is of a size of 10 μ m * 10 μ m; The altitude scale of figure is 10nm.
The AFM shape appearance figure of the bimolecular tunic that Fig. 9 makes for embodiment 7.Wherein appearance is of a size of 10 μ m * 10 μ m; The altitude scale of figure is 10nm.
Embodiment
The solid surface that embodiment 1 spin-coating method prepares three component lipids (DOPC/ESM/Chol) supports adipose membrane.
The chloroformic solution of dioleoyl phospholipid phatidylcholine (DOPC), egg sphingolipid (ESM) and three kinds of phosphatide of cholesterol (Chol) obtains lipid stoste by 2: 2: 1 mixed in molar ratio, and wherein the concentration of lipid stoste is 1.0mg (lipid)/ml (chloroform) solution.With the spin coating instrument at mica surface with the stoste plated film, room temperature places AFM to observe down, Fig. 3.The parameter of spin coating instrument is: rotating speed 1000rpm, and spin coating time 20s, before AFM scanning, sample remains in 45 ℃ the vacuum drying oven.As can be seen from Figure 3, by the supporting film surface irregularity that spin-coating method obtains, film inside is divided into several layers, is unfavorable for the pattern and the inner phase-splitting of observation and analysis film.
The be soaked in water solid surface of the three component lipids (DOPC/ESM/Chol) that spin-coating method makes of embodiment 2 supports adipose membrane.
The adipose membrane that makes in the foregoing description 1 is immersed in the Milli-Q water 12 hours.With after the Milli-Q water flushing, place the AFM liquid pool at room temperature scan Fig. 4.As can be seen, even dry film after the abundant swelling, also can't obtain even curface in water.
The solid surface that embodiment 3 common vesica fusion methods prepare three component lipids (DOPC/ESM/Chol) supports adipose membrane.
Get the stoste among the 2ml embodiment 1, it is prepared into the vesica aaerosol solution of 1.0mg (lipid)/ml (Milli-Q water).Wherein the average-size of vesica is less than 200nm.Get this vesica suspending liquid of 1ml and place the fresh mica surface of peeling off, in 30 ℃ baking oven the insulation 4 hours, use Milli-Q water then carefully, wash away rest solution, and wash repeatedly 10 times, during keep the surface to be in aqueous phase as far as possible.The support membrane that obtains after the flushing is placed the imaging of AFM liquid pool, obtain Fig. 5.As can be seen, the support membrane that obtains among the relative embodiment 1,2, smooth with the film that common vesica fusion method makes, the phase region border is obvious.But the time of fusion of this method cost is long, simultaneously orderly phase region (bright color lump among the figure) shape irregularity.
The solid surface that embodiment 4 usefulness this method prepare three component lipids (DOPC/ESM/Chol) supports adipose membrane.
Get the vesica aaerosol solution that makes among the 60 μ l embodiment 3 and inject the liquid pool inner chamber, load onto injector, wherein the injector first passage connects 50ml Milli-Q water.It is 45 ℃ that the sample stage temperature is made as, and is incubated after 30 minutes to wash with Milli-Q water, is cooled to 30 ℃ then, leaves standstill 10 minutes.Obtain shape appearance figure shown in Figure 6 with afm scan.As can be seen, with the support membrane surfacing that the present invention makes, phase region is clear debates.Simultaneously, the orderly phase region in Fig. 6 all presents circle and ellipse, meets the shape characteristic of DOPC/ESM/Chol three components orderly phase region after the phase-splitting in film.
The solid surface that embodiment 5 usefulness this method prepare two component lipids (POPC/BSM) supports adipose membrane
Get oleoyl phosphatid ylcholine (POPC) and brain sphingolipid (BSM) vesica PBS solution 100 μ l and inject the liquid pool inner chamber, load onto injector, wherein injector one passage connects 100ml PBS damping fluid, the mol ratio of POPC and BSM is 1: 1, the concentration of vesica aqueous solution is 1.0mg (lipid)/ml (PBS) solution, and the vesica size is less than 150nm.The sample stage temperature is set at 50 ℃, is incubated and uses the PBS damping fluid after 40 minutes, be cooled to room temperature (25-28 ℃) then, left standstill 20 minutes.Obtain shape appearance figure shown in Figure 7 with afm scan.As can be seen, with the same surfacing of two component support membranes that the present invention makes, the phase region border is obvious, the phase region irregularity boundary, in order phase region be connected with each other and size bigger, meet POPC/BSM two components shape characteristic after the phase-splitting in film.
The solid surface that embodiment 6 usefulness this method prepare two component lipids (DOPC/GM1) supports adipose membrane
Get DOPC and gangliosides (GM1) vesica aqueous solution 80 μ l and inject the liquid pool inner chamber, load onto injector, wherein the injector first passage connects 100mL Milli-Q water, the mol ratio of DOPC and GM1 is 4: 1, the concentration of vesica aqueous solution is 2.0mg (lipid)/ml (Milli-Q water) solution, and the vesica size is less than 200nm.It is 55 ℃ that the sample stage temperature is made as, and is incubated after 35 minutes to wash with Milli-Q water, is cooled to 30 ℃ then, leaves standstill 15 minutes.Obtain shape appearance figure shown in Figure 8 with afm scan.As can be seen, more smooth with the same surface of two component support membranes that the present invention makes, the phase region border is obvious.The phase region irregularity boundary, phase region internal table surface evenness is lower than phase region on every side, but phase region height height (being presented as that the phase region color is brighter) meets DOPC/GM1 two components shape characteristic after the phase-splitting in film.
Insertion and the distribution of GM1 on three component lipoid (DOPC/ESM/Chol) support membranes in embodiment 7 liquid phases
On the basis of the support membrane that makes in the foregoing description 5, third channel connection concentration at injector is the GM1 aqueous solution 10ml of 0.1 μ g/ml, open channel valve and main valve, inject the GM1 aqueous solution, make the Milli-Q water in the liquid pool inner chamber be displaced inner chamber, close GM1 aqueous solution channel valve and main valve.Leave standstill, allow the GM1 aqueous solution and three component lipoid (DOPC/ESM/Chol) the support membrane effects of injecting.After 30 minutes, open the channel valve and the main valve of Milli-Q water, inject Milli-Q water, displace the GM1 aqueous solution in inner chamber to inner chamber.And then scanning, the AFM shape appearance figure is seen Fig. 9.
Present embodiment is intended to illustrate dirigibility of the present invention and application prospect widely:
(1) because whole operation does not relate to the displacement of AFM instrument and needle point, therefore Fig. 7 and Fig. 5 correspondence is the same area, promptly by the sampling device among the present invention, the present invention has realized in-situ observation, and this dynamics research for the film pattern has crucial meaning.And common vesica fusion method is to be implemented in the variation of home position observation film and process thereof.
(2) there is light tone projection (as shown by arrows) many places among Fig. 7, show that the GM1 molecule has entered supporting film.This is because the GM1 molecule is bigger, in case after inserting supporting film, with projecting membrane surface.Further contrast can find that these projections mainly concentrate on the phase region border, has shown the distribution situation of GM1 molecule in multi-component membranes.Promptly can realize that by the present invention original position changes the residing physical environment of supporting film, thereby change the pattern of supporting film; Under the prerequisite that does not need sample preparation again, increased the component of supporting film, and observed new component region.Common vesica merges rule and requires from the just sample preparation again of preparation vesica solution, cost plenty of time.
Claims (9)
1. method at liquid phase made acid-stable in situ lipid bilayer membrane, it is characterized in that, adopt in-situ high temperature vesica fusion method, in conjunction with atomic force microscope liquid pool punch block design sampling device, under hot conditions, merge vesica, at liquid phase made acid-stable in situ lipid bilayer membrane.
2. by the described method of claim 1, it is characterized in that, described sampling device comprises controllable temperature sample stage (0), liquid pool punch block (1), liquid pool inner chamber (2), liquid injector (3), thermometer (4), flexible pipe (5) and injector passage (6-8), wherein, described sampling device places on the controllable temperature sample stage (0), and the probe of atomic force microscope is installed in liquid pool punch block (1) bottom, and there are three connectors the side, its right side connector connects liquid injector (3), middle connector connects thermometer (4), and the left side connector connects conventional hoses (5), and liquid injector (3) is provided with injector passage (6-8) and connects different types of liquid or solution.
3. by the described method of claim 2, it is characterized in that described liquid pool inner chamber (2) has 0 type circle on every side.
4. by the described method of claim 1, it is characterized in that described method may further comprise the steps:
1) original position is injected vesica solution: before connecting sampling device, to have on the mica of smooth surface or the controllable temperature sample stage that the cover glass solid places atomic force microscope, liquid pool punch block (1) is placed solid surface, in liquid pool inner chamber (2), inject lipid vesica solution;
2) be installed into sampling device: sampling device is connected with liquid pool punch block (1), closes the injector main valve, connect the syringe that required solution is housed in the passage of injector (6-8), thermometer (4) and flexible pipe (5) are installed simultaneously;
3) high temperature merges: open sample stage temperature control power supply, according to used lipid design temperature in the system, with thermometer (4) when reading reaches design temperature, pick up counting, continue heating vesica solution, on mica or cover glass surface, vesica merges the formation bimolecular film each other;
4) original position is removed unnecessary vesica, obtains smooth bimolecular film.
5. by the described method of claim 4, it is characterized in that in the described step 1), the lipid vesica solution that injects in liquid pool inner chamber (2) is 60-100 μ l, the lipid concentration of lipid vesica solution is 0.5-2.0mg/ml, and the vesica size is less than 300nm.
6. by the described method of claim 4, it is characterized in that described step 2) in, the syringe that the required solution of 20-100ml is housed connected.
7. by the described method of claim 4, it is characterized in that in the described step 3), described range of set temperature is 30-60oC.
8. by the described method of claim 4, it is characterized in that in the described step 3), the time of described lasting heating vesica solution is 30-60 minute.
9. by the described method of claim 4, it is characterized in that, in the described step 4), by closing sample stage temperature control power supply, be cooled to room temperature, open injector main valve and solution channel, the solution of 10-20ml is by liquid pool inner chamber (2), take away the vesica that inner chamber does not merge, waste liquid is derived liquid pool inner chamber (2) by flexible pipe (5).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009101992319A CN102072971A (en) | 2009-11-20 | 2009-11-20 | Method for preparing lipid bilayer membrane in liquid phase in situ |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009101992319A CN102072971A (en) | 2009-11-20 | 2009-11-20 | Method for preparing lipid bilayer membrane in liquid phase in situ |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102072971A true CN102072971A (en) | 2011-05-25 |
Family
ID=44031589
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009101992319A Pending CN102072971A (en) | 2009-11-20 | 2009-11-20 | Method for preparing lipid bilayer membrane in liquid phase in situ |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102072971A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114908358A (en) * | 2022-04-14 | 2022-08-16 | 广州孔确基因科技有限公司 | Preparation method and device of amphiphilic molecular layer |
-
2009
- 2009-11-20 CN CN2009101992319A patent/CN102072971A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114908358A (en) * | 2022-04-14 | 2022-08-16 | 广州孔确基因科技有限公司 | Preparation method and device of amphiphilic molecular layer |
| CN114908358B (en) * | 2022-04-14 | 2024-06-04 | 广州孔确基因科技有限公司 | Preparation method and device of amphiphilic molecular layer |
| CN114908358B8 (en) * | 2022-04-14 | 2024-06-21 | 孔确(成都)科技有限公司 | Preparation method and device of amphiphilic molecular layer |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Kim et al. | Collagen-based brain microvasculature model in vitro using three-dimensional printed template | |
| CN103627635B (en) | Multifunctional micro-fluidic chip for cell migration and invasion assay | |
| Matsunaga et al. | Molding cell beads for rapid construction of macroscopic 3D tissue architecture | |
| JP5042235B2 (en) | Bioreactor for cell and tissue culture | |
| JP2013507922A (en) | Microfluidic device for cell culture | |
| Hakam et al. | Evaluation of fibrin-gelatin hydrogel as biopaper for application in skin bioprinting: An in-vitro study | |
| CN102071139A (en) | Microfluidic chip-based cell three-dimensional co-culture method | |
| CN101704685B (en) | Method and device for preparing lipid bilayer membrane in situ in liquid phase | |
| CN109082406B (en) | Method for constructing three-dimensional brain-like development model based on micro-fluidic chip | |
| Demirel et al. | Surface-induced self-assembly of dipeptides onto nanotextured surfaces | |
| Kim et al. | Organoids for advanced therapeutics and disease models | |
| CN102072971A (en) | Method for preparing lipid bilayer membrane in liquid phase in situ | |
| Teng et al. | Molecular level insights into the dynamic evolution of forward osmosis fouling via thermodynamic modeling and quantum chemistry calculation: effect of protein/polysaccharide ratios | |
| Huang et al. | The architecture of co-culture spheroids regulates tumor invasion within a 3D extracellular matrix | |
| WO2019167951A1 (en) | Culture apparatus, method for constructing two or more types of cell structures, and organ-on-a-chip device | |
| Mughal et al. | Organs‐on‐chips: Trends and challenges in advanced systems integration | |
| Dong et al. | Self-assembly of Fmoc-amino acids in capillary confined space forming a parallel ordered fiber network for application in vascularization | |
| CN106092702A (en) | The application in cell climbing sheet SABC detects of the groupization pen of minimizing antibody consumption | |
| KR102037595B1 (en) | Hydrogel membrane fixed vertically in microfluidic chip and preparation method thereof | |
| CN102746580A (en) | Superhydrophobic polypropylene film and preparation method thereof | |
| JP2008278769A (en) | Method for producing three-dimensional aggregate comprising pancreatic islet cells in vitro | |
| Dupas | Wetting of soluble polymers | |
| CN102850567B (en) | Preparation method of material with oriented growth structure cells | |
| Li et al. | Biomaterial scaffolds with biomimetic fluidic channels for hepatocyte culture | |
| CN106486609B (en) | A kind of method that liquid phase prepares ultra-thin two-dimension electrical functionality film |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20110525 |