Modifying glass slide for fixing biomacromolecules and method thereof

Abstract

The invention discloses a modifying glass slide for fixing biomacromolecules and a method thereof. The modifying glass slide comprises a glass slide and a cover glass, wherein a modifying layer which is connected with the surface of the glass slide by using a covalent bond is arranged between the glass slide and the cover glass; a liquid adding hole is formed in the glass slide; the modifying layer is bovine serum albumin marked with biotin or a polyethylene glycol derivative marked with the biotin; the glass slide is a quartz glass slide; the bovine serum albumin marked with the biotin can be 6-8 biotin molecules per albumin molecule; and the mass ratio of polyethylene glycol derivative marked with the biotin to polyethylene glycol derivative without being marked is between 3 and 10 percent. An intermediate dielectric layer is added by virtue of the liquid adding hole to be incubated at room temperature, and sample solutions are added again so as to realize fixation of biomolecules. By using the modifying glass slide, nucleic acid and protein biomolecules can be conveniently fixed, and biological single-molecule imaging data acquisition is realized.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a method for fixing biomolecules on glass slides during observation and analysis of biomolecules. Background technique [0002] Molecular immobilization is the key to single-molecule technology. Only after the molecules are immobilized can single-molecule signals be collected stably for a long time. Untreated glass slides are slightly negatively charged under neutral conditions, and have non-specific adsorption to positively charged protein molecules. In single-molecule imaging, the density of molecules is required to be low enough, and non-specific binding prevents the analyst from Effectively control molecular density. When the molecule is controlled at a low enough level and the molecule is fixed in the focal plane of the excitation light, single-molecule signals can be collected stably without the sample drifting out of the observation window of the objective lens. When...

AI Technical Summary

Problems solved by technology

When fixing the sample, it is necessary to ensure that the fixation does not affect the molecular function, and the polarity of the fluorescent molecules labeled on the biomacromolecule cannot be affected. The polarized fluorescent small molecule emission will complicate data processing.

However, the non-specific binding of the surface charge of the slide will increase the background noise, and the above two problems cannot be solved.

In addition, it is also possible to immobilize the molecules in the gel without modifying the molecules, but the gel immobilization method is not suitable for the study of molecular interactions due to the problem of molecular diffusion.

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