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A set of fluorescent probes for simultaneous visualization of nuclear architecture and cell integrity in living cells

A fluorescent probe and simultaneous display technology, applied in the field of fluorescent probes, can solve the problems of fluorescent reagents limiting biological research, disease diagnosis, pathological research and drug development, etc., to achieve strong photostability, good counterstaining compatibility, strong color effect

Active Publication Date: 2019-11-19
SHANDONG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The lack of such fluorescent reagents has limited biological research, disease diagnosis, pathological research and drug development, and urgently needs to be solved

Method used

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  • A set of fluorescent probes for simultaneous visualization of nuclear architecture and cell integrity in living cells
  • A set of fluorescent probes for simultaneous visualization of nuclear architecture and cell integrity in living cells
  • A set of fluorescent probes for simultaneous visualization of nuclear architecture and cell integrity in living cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Probe preparation.

[0033] 2-(4-ethoxyphenyl)-6-(4-methyl-1-piperazinyl)-1H,3H-2,5-dibenzimidazole-trihydrochloride, 1,1'-bis Octadecyl-3,3,3',3'-tetramethylindolocarbocyanine perchlorate was purchased from Molecular Probes Inc., and the article numbers were Hoechst33342 and DiI, respectively.

[0034] (E)-1-(2-hydroxyethyl)-4-(2-(1-methyl-1H-pyrrole-2-)vinyl)pyridinium iodide salt according to the patent “a kind of Synthesized by the experimental process described in "Pyrrole pyridinium salt fluorescent probe for RNA and nucleolus imaging in living cells (application number: 201310215970.9)".

Embodiment 2

[0035] Example 2: SiHa cell culture.

[0036] Adherently culture SiHa cells in culture medium containing 10% fetal bovine serum at 37°C, 5% CO 2 Cultured in a saturated humidity incubator, and subcultured once every 2-3 days. When the cells grow to the logarithmic phase, culture the slices: ① Soak the coverslips in absolute ethanol for 30 minutes, dry them with an alcohol lamp and place them in a disposable 35mm culture dish for later use; Wash the cells three times with PBS, digest with 1mL 0.25% trypsin for 3-5 minutes, pour out the trypsin carefully, add fresh culture medium and pipette evenly, and count the cells. The concentration is 1×10 5 , and then inoculated into the above-mentioned petri dishes containing coverslips, and cultured in a 5% CO2 incubator to allow the cells to grow on the sheets. After the SiHa cells grow on the slide and cover the glass, it is used for the experiment.

Embodiment 3

[0037] Example 3: (E)-1-(2-hydroxyethyl)-4-(2-(1-methyl-1H-pyrrole-2-)vinyl)pyridinium iodide salt, 2-(4-ethoxy Phenyl)-6-(4-methyl-1-piperazinyl)-1H,3H-2,5-dibenzimidazole-trihydrochloride, 1,1'-octadecyl-3, 3,3',3'-Tetramethyldocarbocyanine perchlorate cell counterstaining method:

[0038] First prepare 5mM (E)-1-(2-hydroxyethyl)-4-(2-(1-methyl-1H-pyrrole-2-)vinyl)pyridinium iodide DMSO solution, 1mM 2-( 4-ethoxyphenyl)-6-(4-methyl-1-piperazinyl)-1H,3H-2,5-dibenzimidazole-trihydrochloride DMSO solution, 1mM 1,1'- Octacosyl-3,3,3',3'-tetramethyldocarbocyanine perchlorate DMSO solution was used as the mother liquor.

[0039] Take 4 μL of (E)-1-(2-hydroxyethyl)-4-(2-(1-methyl-1H-pyrrole-2-)vinyl)pyridinium iodide mother solution, 5 μL of 2-(4- Ethoxyphenyl)-6-(4-methyl-1-piperazinyl)-1H,3H-2,5-dibenzimidazole-trihydrochloride stock solution and 5 μL of 1,1'-octacos Alkyl-3,3,3',3'-tetramethyldocarbocyanine perchlorate mother solution was added to 1 mL of PBS buffer and mixe...

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Abstract

The invention discloses a group of fluorescent probes for simultaneously displaying cell nucleus structure and cell integral morphology in live cells. The group of fluorescent probes comprises a generalized RNA (ribonucleic acid) fluorescent probe with a chemical structure shown in a formula (I), a cell nucleus fluorescent probe with a chemical structure shown in a formula (II), and a cell membrane fluorescent probe with a chemical structure shown in a formula (III). The invention also discloses application of the group of fluorescent probes for labeling or displaying cell integral morphology, especially labeling or displaying the cell nucleus structure including nucleolus distribution. Proofed by experiment, the group of fluorescent probes has the characteristics that the application range is broad, the membrane permeability is good, the cell toxicity is low, and the cell nucleus, cell nucleolus, cell plasma and cell plasma member can be simultaneously imaged; the application prospect is broad.

Description

technical field [0001] The invention relates to a group of fluorescent probes for distinguishing the morphology of living cells, in particular to a group of fluorescent probes for simultaneously displaying the nucleus structure and the overall shape of cells in living cells. Background technique [0002] Nuclear architecture, the spatial distribution of chromosomes and other nuclear components, provides the basic framework for programming and regulating the various functional processes that take place within the nucleus. A large number of studies have found that, compared with normal cells, the structure of the nucleus of cancer cells is completely different. Moreover, among the various morphological changes of tumor cells, the changes in the structure of the nucleus have the most medical diagnostic value. For example, Beale, L.S found and observed changes in the size and shape of nuclei in pharyngeal carcinoma cells exfoliated by patients. [0003] Recently, changes in nu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64C09K11/06
CPCC09K11/06C09K2211/1007C09K2211/1029C09K2211/1044G01N21/6486
Inventor 于晓强潘文胜何秀全田明刚
Owner SHANDONG UNIV
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