A kind of purposes of two-photon fluorescent probe

A two-photon fluorescence and probe technology, applied in the direction of fluorescence/phosphorescence, luminescent materials, material excitation analysis, etc., can solve the problems of low fluorescence quantum yield, short fluorescence lifetime, weak affinity, etc., and achieve low interference of fluorescence, The effect of small molecular volume and large absorption cross section

Active Publication Date: 2021-04-30
ZUNYI NORMAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the probe in Document 1 is a pyridinium salt compound, which is an ionic compound, has strong polarity, poor compatibility with weakly polar lipid rafts, and weak affinity; the probe in Document 1 is easy to combine with nucleic acid and used as a nucleic acid probe [see literature : (1) LiuY, Meng F F, He L W, Yu X Q, Lin W Y. Chem.communications, 2016, 52(57): 8838—8841; (2) Song G F, Miao F, Sun Y M, Yu X Q, Sun J Z, Wong W Y.Sensors and Actuators B Chemical, 2012,173(10):329-337], therefore susceptible to nucleic acid interference, resulting in false positives; and the fluorescence quantum yield of the document 1 probe is quite low (although the patent is not publish this parameter), the fluorescence lifetime of literature 1 probe is relatively short, generally less than 200ps (1ps=10 -12 s) [See literature: (1) Wang Xiaomei et al., Chinese Science (Series E), 32(1): 20-27; (2) Lei Hong et al., Physics, 2003, 32(1): 19-26]

Method used

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  • A kind of purposes of two-photon fluorescent probe
  • A kind of purposes of two-photon fluorescent probe
  • A kind of purposes of two-photon fluorescent probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1 Molecular structure and synthetic route

[0038] A two-photon fluorescent labeling probe DLR has the following molecular structure:

[0039]

[0040] The synthetic route is:

[0041]

[0042] Wherein, intermediates 2, 3, 4, 5, and 6 were all synthesized according to the following documents:

[0043] [1] H. Huang, Q. He, H. Lin, F. Bai, Z. Sun and Q. Li, Polym. Adv. Technol., 2004, 15:84—88

[0044] [2]Huang C., Fan J., Peng X., Lin Z., Guo B., Ren A., Cui J., Sun S., J.Photochem.Photobio.A:Chem.,2008,199( 2–3):144–149

Embodiment 2

[0045] Example 2 Research on the detection process of two-photon fluorescent lipid raft probes

[0046] 2.1 Reagents and instruments

[0047] Dioleoylphosphatidylcholine (DOPC, Xi’an Ruixi Biotechnology Co., Ltd.), dipalmitoylphosphatidylcholine (DPPC, Xi’an Ruixi Biotechnology Co., Ltd.), cholesterol (cholesterol, Xi’an Ruixi Biotechnology Co., Ltd.) , sphingomyelin (sphingomyelin, Xi'an Ruixi Biotechnology Co., Ltd.), methyl-β-cyclodextrin (MβCD, Zibo Qianhui Biotechnology Co., Ltd.]).

[0048] Mira 900-F mode-locked femtosecond titanium sapphire laser (Coherent, USA)

[0049] 2.2 Experimental process

[0050] 2.1 The preparation of the two-photon fluorescent probe DLR was prepared according to the patent application CN2018104756764.

[0051] 2.2 Detection of lipid raft content in cells

[0052] When used to detect the content of lipid rafts: dissolve DLR in the working solution, then incubate the cells with the working solution containing DLR, take out the cells and obs...

Embodiment 3

[0055] Example 3 Research on the Content of Lipid Rafts by Two-photon Fluorescent Probe

[0056] 1) The culture of mouse fibroblasts was prepared according to literature (Huang C., Qu J., Qi J., Yan M., Xu G., Org. Lett., 2011, 13(6), 1462-1465) :

[0057] 2) Preparation of working solution: prepared according to the volume ratio of chloroform and methanol as 9:1;

[0058] 3) Dissolve DLR in the working solution, then incubate the mouse fibroblasts with the working solution containing DLR, take out the cells and observe the fluorescence intensity under the microscope, the cell area with higher fluorescence intensity has higher lipid raft content;

[0059] 4) After adding methyl-β-cyclodextrin (MβCD), incubate mouse fibroblasts, take out the cells and observe that the fluorescence intensity becomes weaker under the microscope, indicating that MβCD can destroy the lipid rafts in the cells, and the lipid raft content is significant decrease;

[0060] 5) After adding 50 μmol / L ...

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Abstract

The present invention relates to the technical field of two-photon fluorescent probes, in particular, to the use of a two-photon fluorescent probe, which is used in cell and tissue imaging to detect lipid raft content and distribution dynamics. The probe can improve the accuracy, precision, sensitivity, and imaging resolution of lipid raft detection, and provide new ideas for further prevention and control of neurodegenerative diseases such as Alzheimer's disease and prion disease.

Description

technical field [0001] The invention relates to the technical field of two-photon fluorescent probes, in particular to the application of a two-photon fluorescent probe. Background technique [0002] Lipid raft (lipid raft) (membrane sheet of 10-100nm) is a microdomain rich in sphingolipids (glycosphingolipids and sphingomyelin) and cholesterol in the plasma membrane, which is involved in cellular processes such as cell transduction and protein transport. Neurodegenerative diseases such as Haimer's disease and prion diseases are closely related. Specifically, lipid rafts are rigid membranes (ordered liquid phase: liquid-ordered phase, lo), floating in a sea of ​​glycerophospholipids (disordered liquid phase: liquid-disordered phase, ld). Its structural characteristics and components are suitable for the interaction and conformational transformation between proteins, and participate in many cellular processes, such as signal transduction, pathogen invasion, cholesterol homeo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64C09K11/06C07D209/86
Inventor 黄池宝
Owner ZUNYI NORMAL COLLEGE
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