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Preparation method of fluorescent carbon dot capable of imaging RNA in living cell for long time in targeting manner and product and application of preparation method

A technology of targeted imaging and fluorescent carbon dots, applied in the field of nanomaterials, to achieve the effect of low biological toxicity, good resistance to photobleaching and biocompatibility

Inactive Publication Date: 2018-01-19
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, carbon dots that can be used to target intracellular RNA have not yet been reported

Method used

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  • Preparation method of fluorescent carbon dot capable of imaging RNA in living cell for long time in targeting manner and product and application of preparation method
  • Preparation method of fluorescent carbon dot capable of imaging RNA in living cell for long time in targeting manner and product and application of preparation method
  • Preparation method of fluorescent carbon dot capable of imaging RNA in living cell for long time in targeting manner and product and application of preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1. Preparation of a green fluorescent carbon dot with strong photobleaching resistance, high biocompatibility, and ability to target and image RNA in living cells for a long time

[0036] A method for preparing green fluorescent carbon dots with strong photobleaching resistance, high biocompatibility, and the ability to target and image RNA in living cells for a long time, comprising the following steps:

[0037] 1) Wash the 25mL tetrafluoroethylene reaction kettle with ultrapure water for use;

[0038] 2) Mix 100mg of m-phenylenediamine, 0.2mL of triethylenetetramine and 4.8mL of ultrapure water into the reaction kettle;

[0039] 3) Place the reaction kettle in an electric heating constant temperature blast drying oven, react at 180°C for 20 hours, and cool to obtain a brownish-yellow liquid;

[0040] 4) neutralize the reaction product with hydrochloric acid solution to pH=7.0;

[0041]5) Filter the neutralized solution with a 0.22 micron filter head, then di...

Embodiment 2

[0046] Example 2. Characterization of the prepared green fluorescent carbon dots with strong photobleaching resistance, high biocompatibility, and ability to target and image RNA in living cells for a long time and their optical properties.

[0047] FTIR-8400S Fourier Transform Infrared Spectrometer (Hitachi, Japan); High Power Transmission Electron Microscope (Tecnai G2F20 S-TWIN microscopy); ESCALAB250 X-ray photoelectron spectrometer; UV Vis Spectrophotometer (Shimadzu, Japan); F-2500 Fluorescence Spectrophotometer (Hitachi, Japan); Absolute PLQuantum yield Spectrum C11347 (HAMAMATSU, Japan); Multi-function microplate reader (BioTek, USA).

[0048] (1) Morphological characterization: take a small amount of prepared carbon dots and dissolve them in water, then drop them on the special copper grid for transmission electron microscope, and observe their morphology with high-power transmission electron microscope after drying. figure 1 shown. It can be seen through a high-magn...

Embodiment 3

[0054] Example 3. Investigation of photostability, cytotoxicity and long-term intracellular RNA targeting imaging ability of prepared carbon dots

[0055] Instruments: F-2500 fluorescence spectrophotometer (Hitachi, Japan); Olympus spinning disk confocal microscopy imaging system (Olympus, Japan); multi-functional microplate reader (BioTek, USA).

[0056] The carbon point that embodiment 1 is made detects, and the result is as follows:

[0057] (1) Photostability investigation: Use F-2500 fluorescence spectrophotometer to measure the fluorescence of carbon dots in different concentrations of sodium chloride solution, hydrogen peroxide solution, BR buffer with different pH gradients and after different UV treatment times Intensity, the result is shown in Figure 4. The results show that the carbon dots prepared by the invention have strong photostability and photobleaching resistance.

[0058] (2) Cell culture: the selected cells are human respiratory epithelial cells (HEp2) i...

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Abstract

The invention relates to a preparation method of a fluorescent carbon dot capable of imaging RNA in a living cell for a long time in a targeting manner and a product and application of the preparationmethod, the fluorescent carbon dot is hydro-thermally synthesized from m-phenylenediamine, an amine compound and water as the raw materials; the synthesis method is simple, the conditions are controllable, the prepared carbon dot has the performances of strong green fluorescence emission, ultralow biotoxicity and strong photobleaching resistance, can image the RNA in the cell for a long time, canserve as an RNA probe to distribute and locate the RNA in the conventional cell, or is applied to the dynamic change of the RNA in the cell, and can further be used to indicate the state of the celland screen an anti-cancer drug taking RNA polymerase I as a target spot.

Description

technical field [0001] The invention belongs to the field of nanomaterials, and in particular relates to a preparation method of strong photobleaching resistance, high biocompatibility, and long-term targeted imaging of RNA fluorescent carbon dots in living cells, and also relates to products and products prepared by the method. application. Background technique [0002] RNA has extremely complex functions in cells, such as transport (tRNA), translation of genetic information (mRNA), molecular machine scaffolding (rRNA), regulation of gene expression levels (miRNA), and catalytic function (ribozymes). Therefore, long-term real-time imaging to monitor the dynamic level of intracellular RNA and its temporal and spatial distribution is of great significance for understanding the physiological function of RNA in cellular activities, the physiological process of RNA-related diseases, and screening anticancer drugs targeting RNA polymerase. Significance. [0003] At present, mor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C01B32/15C09K11/65B82Y20/00B82Y40/00G01N21/64
Inventor 黄承志程云英
Owner SOUTHWEST UNIVERSITY
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