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Method for marking canine kidney cells through chitosan-quantum dot fluorescent probe

A technology of dog kidney cells and fluorescent probes, applied in fluorescence/phosphorescence, material excitation analysis, etc., to achieve the effects of easy operation, controllable fluorescence intensity and color, and good biocompatibility

Inactive Publication Date: 2011-02-23
武汉市疾病预防控制中心 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as a nanomaterial, due to its own physical and chemical properties and the influence of environmental factors, quantum dots also show certain biological toxicity effects in the application process. How to improve the stability and biocompatibility of quantum dot fluorescent probes Sex is an urgent issue

Method used

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  • Method for marking canine kidney cells through chitosan-quantum dot fluorescent probe
  • Method for marking canine kidney cells through chitosan-quantum dot fluorescent probe
  • Method for marking canine kidney cells through chitosan-quantum dot fluorescent probe

Examples

Experimental program
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Effect test

Embodiment 1

[0032] A method for labeling dog kidney cells with a chitosan-quantum dot fluorescent probe, comprising the following steps:

[0033] 1. Prepare 0.0005mol / L green and red carboxymethyl chitosan-CdTe quantum dot solutions:

[0034] Pipette 2.5mL of 0.002mol / L green and red carboxymethyl chitosan-CdTe quantum dot solutions into sterilized 10mL volumetric flasks, add high-purity water (resistivity greater than 18 MΩ·cm) to volume ,spare. The selected chitosan-quantum dot fluorescent probe is formed by self-assembly of carboxymethyl chitosan and CdTe quantum dots in the aqueous phase through electrostatic complexation, covalent binding, chelation, etc., and has good water solubility. and biocompatibility; (carboxymethyl chitosan-CdTe quantum dots are self-made, and the brief preparation process is as follows: CdCl 2 The solution and the thioglycolic acid solution were mixed evenly, and the pH value was adjusted with NaOH solution to be alkaline, and then newly prepared KTeH was ...

Embodiment 2

[0042] A method for labeling dog kidney cells with a chitosan-quantum dot fluorescent probe, the steps are as follows:

[0043] 1. Prepare 0.0001mol / L carboxymethyl chitosan-CdTe quantum dot solution:

[0044] Pipette 0.5mL of 0.002mol / L yellow-green carboxymethyl chitosan-CdTe quantum dot solution into a sterilized 10mL volumetric flask, add high-purity water (resistivity greater than 18 MΩ·cm) to volume, and set aside . The selected chitosan-quantum dot fluorescent probe is formed by self-assembly of carboxymethyl chitosan and CdTe quantum dots in the aqueous phase through electrostatic complexation, covalent binding, chelation, etc., and has good water solubility. and biocompatibility;

[0045] 2. Culture of dog kidney cells:

[0046] The dog kidney cells (MDCK) that were full and in good growth condition in the T25 cell culture flask were digested with 0.05% EDTA-trypsin into a single cell suspension, and 1 × 10 cells were taken after counting the cells. 5 The dog kidn...

Embodiment 3

[0053] A method for labeling dog kidney cells with a chitosan-quantum dot fluorescent probe, the steps are as follows:

[0054] 1. Prepare 0, 0.00005, 0.0001, 0.00025, 0.0005, 0.001mol / L carboxymethyl chitosan-CdTe quantum dot solution:

[0055] Pipette 0, 0.25, 0.5, 1.25, 2.5, 5mL of 0.002mol / L yellow-green carboxymethyl chitosan-CdTe quantum dot solution into a sterilized 10mL volumetric flask, add high-purity water (resistivity greater than 18 MΩ·cm) constant volume, spare. The selected chitosan-quantum dot fluorescent probe is formed by self-assembly of carboxymethyl chitosan and CdTe quantum dots in the aqueous phase through electrostatic complexation, covalent binding, chelation, etc., and has good water solubility. and biocompatibility;

[0056] 2. Culture of dog kidney cells:

[0057] The dog kidney cells (MDCK) that were full and in good growth condition in the T25 cell culture flask were digested with 0.05% EDTA-trypsin into a single cell suspension, and 1 × 10 ce...

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Abstract

The invention discloses a method for marking canine kidney cells through a chitosan-quantum dot fluorescent probe. The method comprises the following steps of: A, preparing solution of a carboxymethyl chitosan-CdTe quantum dot; transferring the solution of the carboxymethyl chitosan-CdTe quantum dot to a sterilized volumetric flask by using a pipette according to the concentration of the carboxymethyl chitosan-CdTe quantum dot, and adding high-purity water for constant volume; B, digesting the canine kidney cells overgrown in a T25 cell culture flask by using ethylene diamine tetraacetic acid (EDTA)-trypsin so as to form unicellular suspension, counting cells, inoculating the canine kidney cells to a cell culture dish, and adding nutrient solution for culturing; and C, after the canine kidney cells grow adherently, adding the chitosan-quantum dot fluorescent probe into the culture dish, and incubating the probe and the canine kidney cells together to obtain canine kidney cells marked with fluorescence. The method is easy and convenient to operate, has high fluorescent efficiency, good biocompatibility and long stable time, and can be used for observing the cells for a long time; and the fluorescent probe does not have cytotoxicity.

Description

technical field [0001] The invention belongs to the technical field of analysis and detection of nanomaterials for labeling organisms and biological systems, and more particularly relates to a method for labeling canine kidney cells with a chitosan-quantum dot fluorescent probe. The markers can be used to study the physiological activities of cells, as well as the transport pathways of various receptors inside and outside cells and the study of the interaction between exogenous pollutants and cells. Background technique [0002] Chitosan is the product of deacetylation from chitin, and its macromolecular chain has many hydroxyl and amino groups distributed. In addition to the advantages of multifunctionality, biocompatibility, biodegradability, and low cytotoxicity, chitosan also has a unique ability to cross cell membranes. However, because chitosan is only soluble in dilute acid and some specific solvents, its application is greatly limited. Carboxymethyl chitosan is a c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 何振宇周培疆朱洪浩
Owner 武汉市疾病预防控制中心
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