A Fluorescence Detection Method of Amphiphilic Monolayer Molecular Film

A fluorescence detection and molecular film technology, applied in the field of fluorescence detection, can solve the problems of inaccurate observation structure, damaged sample surface, non-reusable, etc., and achieve the effect of clear results, low cost and saving detection cost

Active Publication Date: 2019-10-29
SHAANXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, after the fluorescence observation of the prepared single-layer film, the residue of oil droplets caused some influence on the further observation of the atomic force microscope and other instruments, which damaged the surface of the sample, resulting in inaccurate observation of the structure, and the cover glass Tablets are also damaged and cannot be reused

Method used

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  • A Fluorescence Detection Method of Amphiphilic Monolayer Molecular Film
  • A Fluorescence Detection Method of Amphiphilic Monolayer Molecular Film
  • A Fluorescence Detection Method of Amphiphilic Monolayer Molecular Film

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] S1: Preparation of amphiphilic monolayer molecular membranes by conventional membrane production methods;

[0028] In this example, LB technology was used to prepare a mixed monolayer membrane of sphingomyelin / 1 palmitic acid-2 oleic acid-phosphatidylcholine / cholesterol (hereinafter referred to as BSM / POPC / Chol).

[0029] Dissolve BSM / POPC / Chol (2:2:1, mol:mol:mol) in chloroform and methanol (10:1, V:V) to form a mixed lipid solution with a concentration of 0.01mmol / L. In KSV-Minitrough Prepare in the instrument (Finland KSV company, tank: 323mm×75mm). Use a micro-injector to drop 50uL of the mixed lipid solution onto the surface of pure water and let it spread automatically. After 30 minutes, wait for the organic solvent on the liquid surface to completely volatilize , The film was pressed at a speed of 10mm / min to obtain a BSM / POPC / Chol mixed monolayer molecular film with a thickness of 3nm.

[0030] S2: Under a fixed film pressure (1mN / m~55mN / m), the monolayer is transferr...

Embodiment 2

[0035] S1: Preparation of amphiphilic monolayer molecular membranes by conventional membrane production methods;

[0036] The sphingomyelin and 1 palmitic acid-2 oleic acid-phosphatidylcholine (hereinafter referred to as BSM / POPC) were mixed in a molar ratio of 1:1 and dissolved in chloroform and methanol (3:1, V:V). ) Was formed into a mixed lipid solution with a concentration of 0.5mmol / L, prepared in the KSV-Minitrough instrument (KSV, Finland, tank: 323mm×75mm), and 20uL of the mixed lipid solution was added dropwise to pure water with a micro-injector The surface is allowed to spread automatically. After 15 minutes, after the organic solvent on the liquid surface is completely volatilized, the film is pressed at a speed of 10 mm / min to obtain a BSM / POPC hybrid monolayer molecular film with a thickness of 3 nm.

[0037] S2: Under a fixed film pressure (1mN / m~55mN / m), the monolayer is transferred to the newly dissociated ultra-thin mica substrate using the vertical film method a...

Embodiment 3

[0042] S1: Preparation of amphiphilic monolayer molecular membranes by conventional membrane production methods;

[0043] Dissolve sterol molecules in chloroform and methanol (10:1, V:V) to form a lipid solution with a concentration of 0.01 mmol / L, and prepare it in a KSV-Minitrough instrument (KSV, Finland, tank: 323mm×75mm) , Use a micro-injector to drop 50uL of the lipid solution onto the surface of the pure water and let it spread automatically. After 30 minutes, after the organic solvent on the surface of the liquid has completely evaporated, press the film at a speed of 20mm / min to obtain a sterol monolayer Molecular membrane.

[0044] S2: Under a fixed film pressure (1mN / m~55mN / m), the monolayer is transferred to the newly dissociated ultra-thin mica substrate with a thickness of 0.01mm using the vertical film method at a speed of 15-25mm / min , The film forming temperature is controlled at 21±0.5℃;

[0045] S3: Drop a drop of water on the cleaned cover glass with a thickness...

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Abstract

The invention belongs to the technical field of fluorescence detection and particularly relates to a fluorescence detection method for an amphiphilic single-layer molecular film. A single-layer film transferred to an ultrathin mica sheet is adsorbed to cover glass containing water in surface through film making, film formation employing a vertical film lifting method and fluorescence detection after dripping adsorption, the surface tension of the water and the transparency are skillfully utilized to nondestructively test the molecular film on the ultrathin mica sheet, the damage to the film is avoided, the undamaged film is ensured, further operations such as atomic force microscope observation are facilitated and the accurate detection result is ensured. The mica sheet can be easily separated from the cover glass through tweezers after fluorescence microscope detection is completed and the cleaned cover glass can still be repeatedly used, so that the detection cost is reduced. Furthermore, the detection method is clear in detection result, relatively low in cost, nontoxic, free of pollution and suitable for large-scale popularization and application.

Description

Technical field [0001] The invention belongs to the technical field of fluorescence detection, and specifically relates to a fluorescence detection method of a single-layer molecular film, particularly a fluorescence detection method of an amphipathic single-layer molecular film. Background technique [0002] Monolayer lipid membrane is widely used as a model of biological membrane. There are many components of biological membranes. For the convenience of research, various artificial lipid membrane structures composed of single or several lipids are often used. Monolayer membrane is one of them. At the same time, it is also possible to embed proteins to form a reconstructed membrane. The artificial membrane can be used to conduct in-depth research on various physical and chemical properties of the membrane. The study of monomolecular lipid membranes has a wide range of potential applications in the fields of biomedical research and biosensor development. [0003] At present, for ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
CPCG01N21/6458
Inventor 郝长春张蕾徐国庆孙润广
Owner SHAANXI NORMAL UNIV
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