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Fluorescence labeling reagent kit for cell nucleus and labeling method of reagent kit

A technology for fluorescent labeling and cell nuclei, applied in biochemical equipment and methods, measuring devices, instruments, etc., can solve the problems of complex steps, low efficiency, and high cost, and achieve the effect of simple method, convenient operation, and strong practicability

Active Publication Date: 2018-01-12
王艳丽
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] On the other hand, there is currently no research that uses the unique chemical advantages of quantum dots to directly achieve selective localization of the nucleus. Previous studies have focused on cross-linking quantum dots with guiding molecules for nuclear localization, such as antibodies and nuclear localization signals. Only in this way can the positioning and marking of the nucleus by quantum dots be realized, which leads to complicated steps of quantum dots for fluorescent labeling of the nucleus, high cost and low efficiency
Moreover, only GQDs and nuclear markers such as DAPI or PI are used in combination to mark the nucleus at home and abroad, and there is no research that directly uses the unique optical properties of GQDs to demonstrate long-term stability of the nucleus without any special targeting molecules. track and mark

Method used

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  • Fluorescence labeling reagent kit for cell nucleus and labeling method of reagent kit
  • Fluorescence labeling reagent kit for cell nucleus and labeling method of reagent kit
  • Fluorescence labeling reagent kit for cell nucleus and labeling method of reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] GQDs and DAPI fluorescence stability comparison experiment:

[0031] To study the photostability of GQDs, we used DAPI, a traditional nuclear fluorescent dye, as a reference. details as follows:

[0032] 1. 4T1 cells (mouse breast cancer cells) were cultured in DMEM medium containing 10% fetal bovine serum at 37 o C with 5% CO 2 cultivated under conditions.

[0033] 2. Wash the cells twice with PBS solution, 2-5 minutes each time.

[0034] 3. Cells were fixed with 4% paraformaldehyde-PBS for 15 min.

[0035] 4. Wash the cells twice with PBS solution, 2-5 minutes each time.

[0036] 5. Divide the treated cells into two groups. GQDs (10 mg / L) and DAPI (2 mg / L) were added dropwise, and incubated at room temperature for 5 min.

[0037] 6. Wash the cells twice with PBS solution.

[0038] 7. Add mounting solution dropwise to mount the slides.

[0039] 8. Place the cells under a confocal microscope, excite with 405 nm excitation light, and observe the cell nucleus ima...

Embodiment 2

[0042] GQDs efficiently target the nucleus of different cells:

[0043] To investigate the performance of GQDs on the nuclei of different cell types, we performed experiments using several different cell types. details as follows:

[0044]1. 4T1 cells, C17.2 cells (mouse neural stem cells), GES-1 cells (human gastric mucosal cells) were cultured in DMEM or RPMI1640 medium containing 10% fetal bovine serum, at 37 o C with 5% CO 2 cultivated under conditions.

[0045] 2. Wash the cells twice with PBS solution, 2-5 minutes each time.

[0046] 3. Cells were fixed with 4% paraformaldehyde-PBS for 15 min.

[0047] 4. Wash the cells twice with PBS solution, 2-5 minutes each time.

[0048] 5. Add GQDs (10 mg / L) dropwise to the treated cells and incubate at room temperature for 5 min.

[0049] 6. Wash the cells twice with PBS solution.

[0050] 7. Add mounting solution dropwise to mount the slides.

[0051] 8. Place the cells under a confocal microscope, excite with 405 nm exci...

Embodiment 3

[0054] GQDs efficiently target the nuclei of different tumors and normal tissues:

[0055] To study the performance of GQDs on the nuclei of different tumor and normal tissues, we used several different kinds of tumor and normal tissues for experiments. details as follows:

[0056] 1. Use different types of tumor and normal tissue sections, such as liver cancer tissue, breast cancer tissue, cervical cancer tissue, lung cancer tissue, heart, liver, spleen, lung, kidney, brain, etc.

[0057] 2. Tissue sections were fixed with 4% paraformaldehyde-PBS fixative for 15 min.

[0058] 3. Tissue slices were washed twice with PBS solution, 2-5 minutes each time.

[0059] 5. To the treated tissue sections, first drop SYTO17 (5 µM, a nuclear labeling dye, excitation wavelength 605 nm) and incubate at room temperature for 5 min, then add GQDs (10 mg / L) dropwise and incubate at room temperature for 5 min.

[0060] 6. Tissue slices were washed twice with PBS solution.

[0061] 7. Add mou...

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Abstract

The invention relates to a fluorescence labeling reagent kit for a cell nucleus and a labeling method of the reagent kit. Particularly, the invention provides a fluorescence labeling method of a graphene quantum dot for a cell nucleus, and a stable fluorescence labeling reagent kit of the quantum dot for the cell nucleus. The method and the reagent kit is simple and practicable, can perform fluorescence labeling on the cell nucleus efficiently and stably; a stable tracing method of the cell nucleus is provided for a long-time observation study of a cell; and the method and the reagent kit canbe used for the efficient fluorescence labeling of the cell nucleus of the cell, a tissue and the like.

Description

technical field [0001] The invention relates to the technical field of graphene quantum dot bioluminescent probes and cell fluorescence labeling. Specifically, the invention provides a method for directly stably fluorescently labeling cell nuclei with new graphene quantum dots, and containing the above quantum dots for use in Kit for stable fluorescent labeling of nuclei. Background technique [0002] In traditional fluorescent labeling techniques for cell nuclei, organic fluorescent dyes such as 4',6-diamidino-2-phenylindole (DAPI) and internalidine iodide (PI) are mostly used for nuclear labeling. Both DAPI and PI can strongly bind to the chromosomal DNA in the nucleus, so they can be used to localize the nucleus and observe changes in structure and shape. [0003] Currently, with the rapid development of material science, graphene quantum dots (GQDs) are gradually being used in the biomedical field due to their unique optical properties. Compared with organic fluorescen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533C12Q1/02
Inventor 王艳丽姚晨婕丁琳李晨晨章康康吴明红
Owner 王艳丽
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