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Long-time cell membrane imaging agent and preparation method thereof

A cell membrane, long-term technology, applied in chemical instruments and methods, fluorescence/phosphorescence, luminescent materials, etc., can solve the problem of dyeing time limited to 30 minutes to 90 minutes, and achieve excellent imaging effects, increase efficiency, and improve imaging quality. and the effect of imaging time

Active Publication Date: 2015-01-28
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to prolong the effective dyeing time and reduce the endocytosis of dyes, some companies have developed new dye molecules, such as CellMask TM Orang and DeepRed series mainly use negatively charged lipophilic groups as anchors to prolong the staining time of cell membranes, but the staining time is limited to 30 minutes to 90 minutes

Method used

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  • Long-time cell membrane imaging agent and preparation method thereof
  • Long-time cell membrane imaging agent and preparation method thereof
  • Long-time cell membrane imaging agent and preparation method thereof

Examples

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Embodiment 1

[0025] The design, synthesis and cell membrane fluorescent staining methods of long-time cell membrane fluorescence imaging reagents (component A: chitosan-10% cholesterol-10% biotin and component B: avidin-FITC) are as follows:

[0026] Reagent design: The molecular structure diagram of the imaging reagent is shown in figure 1 , the reagent consists of two components: Component A uses glycol chitosan as the backbone, and the side chain contains 10% polyethylene glycol 2000-cholesterol (PEG2000-cholesterol) hydrophobic units and 10% biotin; component B consists of avidin grafted with fluorescein isothiocyanate FITC. The ethylene glycol chitosan macromolecule has good biocompatibility and water solubility. The hydrophobic side chain of cholesterol is connected to the amino group of chitosan through PEG2000 with a molecular weight above 500, that is, NHS-PEG2000-cholesterol, to increase water solubility. The FITC fluorescent molecule is connected to the avidin molecule through...

Embodiment 2

[0034] The implementation method of the present embodiment is consistent with the method in Example 1, except that the ethylene glycol chitosan macromolecule molecular weight of selection is about 10000, and the cholesterol hydrophobic unit in the NHS-PEG2000-cholesterol molecule is changed into phospholipid molecule (1,2) successively -myristoylphosphatidylethanolamine, DMPE), alkanes (eighteen carbon atoms, C18) or vitamin E and other hydrophobic side chains, synthesized to obtain three kinds of cell membrane fluorescence imaging reagents.

Embodiment 3

[0036] In order to realize fluorescence imaging of cell membranes in other colors, the implementation method of this embodiment is similar to the method in Example 1, except that the molecular weight of the selected ethylene glycol chitosan is about 100,000, and the fluorescent molecule FITC in Example 1 is replaced. Become the sulforhodamine B sulfonyl chloride fluorescent reagent, and the implementation method is consistent with the method in Example 1.

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Abstract

The invention discloses a long-time cell membrane imaging agent, which is characterized by comprising a component A and a component B. The component A is a glycol chitosan polymer with the side chain containing biotin and a hydrophobic unit; and the component B is an avidin molecule grafted with fluorescein isothiocyanate FITC. The cell membrane imaging reagent provided by the invention has good biological compatibility and good imaging effect, and can stay on the cell membrane for a long time without being swallowed, and can be used as a novel cell membrane imaging agent.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a long-time cell membrane fluorescence imaging reagent and a preparation method of the reagent. Background technique [0002] The cell membrane not only serves as the boundary of the cell structurally, providing a relatively stable internal environment for the cell, but also plays a vital role in the process of material transportation, energy conversion and information transmission between the cell and the external environment. As a powerful research method, cell membrane imaging technology uses fluorescent dyes to label and trace the cell membrane, revealing many important biological processes including pinocytosis, exocytosis, cell division and apoptosis. [0003] However, some traditional lipophilic dyes will endocytosis quickly, and the imaging time is short, which brings great limitations to the research. For example, the currently commercially available fluorescent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64C09K11/06C08B37/08
Inventor 吴富根贾浩然王宏银
Owner SOUTHEAST UNIV
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