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Fluorescence imaging reagent for cell membrane based on multi-site anchoring and preparation method thereof

A multi-site anchoring, fluorescence imaging technology, applied in fluorescence/phosphorescence, chemical instruments and methods, luminescent materials, etc., can solve problems such as difficult large-scale application, difficult technological breakthrough, cell endocytosis, etc. Improve dyeing efficiency and good biocompatibility

Active Publication Date: 2014-12-24
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since they are all small molecule fluorescent dyes, they are easily endocytized by cells (especially when the staining time is long, the endocytosis is more obvious), which further makes it difficult for this single-site anchoring method to have a technical breakthrough
Coupled with its complex synthesis and high cost, it is difficult to apply on a large scale

Method used

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  • Fluorescence imaging reagent for cell membrane based on multi-site anchoring and preparation method thereof
  • Fluorescence imaging reagent for cell membrane based on multi-site anchoring and preparation method thereof
  • Fluorescence imaging reagent for cell membrane based on multi-site anchoring and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The multi-site anchored cell membrane fluorescence imaging reagent chitosan-30% cholesterol-2% FITC design, synthesis and cell membrane fluorescence staining methods are as follows:

[0028] Reagent design: The molecular structure diagram of the imaging reagent is shown in figure 1 , the reagent is based on glycol chitosan macromolecules, and the side chain contains 30% polyethylene glycol 2000-cholesterol (PEG2000-cholesterol) hydrophobic units and 2% FITC fluorescent units. The ethylene glycol chitosan macromolecule has good biocompatibility and water solubility, and its molecular weight needs to be greater than 10,000. The cholesterol hydrophobic unit is linked to the chitosan amino group through PEG2000, and the PEG chain segment can increase water solubility. The FITC fluorescent molecule is directly linked to the chitosan macromolecule through reaction with amino groups, and the FITC fluorescent molecule realizes fluorescence imaging. The cholesterol hydrophobi...

Embodiment 2

[0033] The implementation method of the present embodiment is consistent with the method in Example 1, except that the high molecular weight of glycol chitosan selected is about 100,000, and the percentage of the cholesterol anchor unit accounting for the chitosan repeating unit number is changed from 30% to 10%. , the composition of the synthetic imaging reagent is chitosan-10% cholesterol-2% FITC, and the imaging effect after staining is shown in Figure 3(b).

Embodiment 3

[0035] The implementation method of the present embodiment is consistent with the method in Example 1, except that the high molecular weight of glycol chitosan selected is about 10000, and the dyeing time is extended from 5min to 1 hour, and the imaging effect after dyeing is shown in Figure 3 (c) Show. It can be seen from the figure that although the staining time was prolonged, it did not cause obvious internalization of the dye.

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Abstract

The invention provides a fluorescence imaging reagent for a cell membrane based on multi-site anchoring. The fluorescence imaging reagent is an ethylene glycol chitosan polymer of which a side chain contains a hydrophobic unit and a fluorescent unit. The fluorescence imaging reagent for the cell membrane is based on a multi-site anchoring technology and has the advantages of good biocompatibility, simplicity for synthesis, low cost and short dyeing time; besides, the fluorescence imaging reagent is not easily intemalized by cells and can be used as a specific tracer labeling probe of the cell membrane.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a cell membrane-specific fluorescence imaging reagent based on multi-site anchoring technology, and also relates to a preparation method of the reagent. Background technique [0002] The cell membrane not only wraps individual cells structurally to form a stable internal environment, but also participates in material transport, energy transfer, signal transduction, vesicle transport, cell growth, differentiation, adhesion, transfer, stress, and endocytosis functionally , exocytosis, apoptosis, necrosis, autophagy and many other important biological processes, so more and more attention has been paid to the research on cell membranes. Fluorescent imaging, which uses fluorescent dyes to label and track cell membranes, is a powerful tool for studying the structure and function of cell membranes. [0003] However, currently available lipophilic fluorescent dyes such as DiD f...

Claims

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Application Information

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IPC IPC(8): C08B37/08C09K11/06G01N21/64
Inventor 吴富根王宏银贾浩然陈战
Owner SOUTHEAST UNIV
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