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Application of fluorescent carbon dots based on N-methyl o-phenylenediamine hydrochloride in lysosome target

A technology of methyl o-phenylenediamine hydrochloride and fluorescent carbon dots, which is applied in the direction of fluorescence/phosphorescence, luminescent materials, preparations for in vivo tests, etc., can solve the problems of long cultivation time, achieve high quantum yield, Good light stability and easy handling

Active Publication Date: 2019-07-26
ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are few reports on the inherent lysosomal targeting of carbon dots, and there are limitations such as long culture time and requirements for post-modification of lysosomal targeting groups

Method used

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  • Application of fluorescent carbon dots based on N-methyl o-phenylenediamine hydrochloride in lysosome target
  • Application of fluorescent carbon dots based on N-methyl o-phenylenediamine hydrochloride in lysosome target
  • Application of fluorescent carbon dots based on N-methyl o-phenylenediamine hydrochloride in lysosome target

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The steps for the synthesis of carbon dots are as follows:

[0037] First, weigh 0.009 g (0.05 mmol) of N-methyl-1,2-phenylenediamine dihydrochloride solid powder, dissolve it in 10 mL of absolute ethanol, and transfer the solution to a high-pressure PTFE lined In the reaction kettle, place the reaction kettle in an oven at 180°C and reflux for 12 h, and then cool the reaction kettle to room temperature to obtain a brown-black solution. Then the crude product was purified by silica gel column chromatography. Using ethyl acetate as the eluent, the solvent was removed under vacuum and further dried to finally obtain purified carbon dots.

Embodiment 2

[0039] Changes in fluorescence intensity of carbon dots in different solvents

[0040] Add 2 mL of different solvents to the cuvette, add 8 µL of carbon spot storage solution, and measure the fluorescence spectrum. The experimental results show that: as shown in Table 1, Table 1 shows the physical data and fluorescence intensity changes of 40μg / mL carbon dots in different solvents under 430nm excitation. It can be seen that the fluorescence of carbon dots in aqueous solution is weak and can be ignored. Regardless, it has the potential of cell imaging without washing.

[0041] Table 1 Physical data and fluorescence intensity changes of carbon dots in different solvents

[0042]

Embodiment 3

[0044] Fast cell imaging study of carbon dots

[0045] In a 37 ℃, 95% air, 5% carbon dioxide incubator, inoculate Hela cells into a special laser confocal culture dish containing 10% fetal bovine serum for culture. After incubating for 24 hours, excitation with an excitation wavelength of 488 nm light source for confocal imaging to observe the time when the carbon dots enter the cell.

[0046] Such as image 3 As shown, the fluorescence response of cells can be observed at an excitation wavelength of 488nm; cell imaging data shows that the carbon dots can quickly enter cells and have good cell permeability.

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Abstract

The invention provides an application of fluorescent carbon dots based on N-methyl o-phenylenediamine hydrochloride in lysosome target. The fluorescent carbon dots are prepared by dissolving the N-methyl o-phenylenediamine hydrochloride into absolute ethyl alcohol; the fluorescent carbon dots can quickly penetrate through cytomembrane and be gathered in lysosome, to be applied to washing-free lysosomal imaging of living cell and living bodies. The carbon dots in the invention are very simple to synthesize and convenient to operate, has fluorophores, and have the advantages of good light stability and high quantum yield; the fluorescent carbon dots can quickly enter cells, and can be naturally targeted at the lysosome; and the fluorescent carbon dots can be applied to quick and washing-freeimage in the living cells and living bodies.

Description

Technical field [0001] The invention relates to the field of preparation of fluorescent nanomaterials, in particular to the application of fluorescent carbon dots based on N-methyl o-phenylenediamine hydrochloride in lysosome targeting. Background technique [0002] Lysosomes are single membrane-bound cytoplasmic acidic organelles found in almost all eukaryotic cells. As a cellular stomach, lysosomes play a key role in many physiological processes and signal transduction pathways, including intracellular transport, membrane circulation, endocytosis, apoptosis, and autophagy. Lysosomal dysfunction is related to neurodegenerative diseases, lipid and glucose metabolism disorders, infectious diseases and various immune system diseases. In addition, lysosomes are essential for macromolecular circulation, bone remodeling, and intracellular transport. More and more researches focus on lysosomal imaging in order to better understand the status and biological role of lysosomes. [0003] ...

Claims

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Application Information

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IPC IPC(8): G01N21/64A61K49/00C09K11/02C09K11/65
CPCA61K49/0019C09K11/025C09K11/65G01N21/6428
Inventor 杨冉孙远强秦皓月李朝辉屈凌波
Owner ZHENGZHOU UNIV
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