A cell membrane fluorescence imaging reagent based on multi-site anchoring and its preparation method

A multi-site anchoring and fluorescence imaging technology, applied in the direction of fluorescence/phosphorescence, chemical instruments and methods, luminescent materials, etc., can solve problems such as difficult large-scale application, difficult technological breakthroughs, and endocytosis, achieving low cost, Improved dyeing efficiency and good biocompatibility

Active Publication Date: 2016-10-12
SOUTHEAST UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • A cell membrane fluorescence imaging reagent based on multi-site anchoring and its preparation method
  • A cell membrane fluorescence imaging reagent based on multi-site anchoring and its preparation method
  • A cell membrane fluorescence imaging reagent based on multi-site anchoring and its preparation method

Examples

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Effect test

Embodiment 1

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

[0028] Reagent design: see the molecular structure diagram of the imaging reagent figure 1 , the reagent uses glycolchitosan polymer as the backbone, and the side chain contains 30% polyethylene glycol 2000-cholesterol (PEG2000-cholesterol) hydrophobic unit and 2% FITC fluorescent unit. The ethylene glycol chitosan polymer 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 amino group of chitosan through PEG2000, and the PEG segment can increase the water solubility. The FITC fluorescent molecule is directly linked to the chitosan polymer through the reaction with the amino group, and the FITC fluorescent molecule realizes fluorescence imaging. Cholesterol hydrophobic units can be...

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 cell membrane fluorescence imaging reagent based on multi-site anchoring, which is an ethylene glycol chitosan macromolecule whose side chain contains a hydrophobic unit and a fluorescent unit. The cell membrane fluorescent imaging reagent is based on multi-site anchoring technology, has good biocompatibility, simple synthesis, low cost, short staining time and is not easy to be endocytized by cells, and can be used as a cell membrane-specific tracer labeling probe.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a cell membrane-specific fluorescent 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 structurally wraps individual cells to form a stable internal environment, but also functionally participates in material transport, energy transfer, signal transduction, vesicle transport, cell growth, differentiation, adhesion, transfer, stress, endocytosis , exocytosis, apoptosis, necrosis, autophagy and many other important biological processes, so the research on the cell membrane has received more and more attention. Fluorescence imaging technology, which uses fluorescent dyes to label and track cell membranes, is a powerful means to study the structure and function of cell membranes. [0003] However, currently commercially available lipophilic fluorescent dy...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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