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Quantum-dot-based method for carrying out in-situ and real-time detection on heavy metal ions in cells

A technology for real-time detection of heavy metal ions, used in material excitation analysis, fluorescence/phosphorescence, etc., can solve problems such as inappropriate real-time detection of mercury and copper ions, inability to describe the distribution and dynamic changes of mercury and copper ions, and achieve fluorescence stabilization time. Long, easy-to-operate effect

Active Publication Date: 2014-12-17
武汉市疾病预防控制中心 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, there are many methods for detecting heavy metal ions such as mercury and copper in cells, such as atomic absorption / emission spectrometry, high performance liquid chromatography, inductively coupled plasma mass spectrometry, electrochemical method, chemiluminescence method, etc., these The method has high sensitivity and selectivity and is often used in quantitative analysis. However, they are not suitable for real-time detection of mercury and copper ions in vivo, and cannot describe the distribution and dynamic changes of mercury and copper ions in cells.

Method used

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  • Quantum-dot-based method for carrying out in-situ and real-time detection on heavy metal ions in cells

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Embodiment 1

[0034] The method for in situ and real-time detection of mercury ions in dog kidney cells based on a quantum dot-labeled cell system comprises the following steps:

[0035] 1. Prepare 0.0005mol / L carboxymethyl chitosan-CdTe quantum dot solution:

[0036] Use a pipette to pipette 2.5 mL of 0.002 mol / L carboxymethyl chitosan-CdTe quantum dot solution into a sterilized 10 mL volumetric flask, add high-purity water (resistivity greater than 18 MΩ·cm) to constant volume, and set aside. The selected carboxymethyl chitosan-CdTe quantum dot fluorescent probe is formed by self-assembly of carboxymethyl chitosan and CdTe quantum dots in the water phase through electrostatic complexation, covalent bonding, chelation, etc. Good water solubility and biocompatibility; (Carboxymethyl chitosan-CdTe quantum dots are self-made, the brief process of preparation is as follows: CdCl 2 Solution and thioglycolic acid solution were mixed evenly, and then the pH value was adjusted with NaOH solution ...

Embodiment 2

[0048] The method for in situ and real-time detection of copper ions in dog kidney cells based on a quantum dot-labeled cell system comprises the following steps:

[0049] 1. Prepare 0.0001mol / L carboxymethyl chitosan-CdTe quantum dot solution:

[0050] Use a pipette to pipette 0.5 mL of 0.002 mol / L carboxymethyl chitosan-CdTe quantum dot solution into a sterilized 10 mL volumetric flask, add high-purity water (resistivity greater than 18 MΩ·cm) to constant volume, and set aside. The selected carboxymethyl chitosan-CdTe quantum dot fluorescent probe is formed by self-assembly of carboxymethyl chitosan and CdTe quantum dots in the water phase through electrostatic complexation, covalent bonding, chelation, etc. Good water solubility and biocompatibility;

[0051] 2. Culture of dog kidney cells:

[0052] The dog kidney cells (MDCK) in the T25 cell culture flask were overgrown and in good growth state were digested with 0.05% volume ratio of EDTA-trypsin into a single-cell susp...

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Abstract

The invention discloses a quantum-dot-based method for carrying out in-situ and real-time detection on heavy metal ions in cells. The method comprises the following steps of: A. preparing a carboxymethyl chitosan-CdTe quantum dot solution with certain concentration; B. digesting canine kidney cells overgrowing in a T25 cell culture flask bottle into a unicellular suspension, inoculating into a cell culture dish, and culturing; C. adding the carboxymethyl chitosan-CdTe quantum dot solution into the culture dish, and hatching for a period of time, thereby obtaining the canine kidney cells marked with fluorescence; and D. adding a DMEM (dulbecco's modified eagle medium) nutrient solution containing heavy metal ions with gradient concentration and unknown concentration respectively in washed cells in the culture dish, hatching together with the cells, detecting the average fluorescent intensity of the cells respectively by a flow cytometry, and drawing a standard curve through the changes of the cell fluorescent intensity, so that the concentration of the heavy metal ions in the cells can be calculated. The method is simple and convenient to operate; the fluorescence of marked cells can be quenched regularly as the increase of heavy metal ions, and the real-time and in-situ detection on the outer source heavy metal ions in the cells is realized.

Description

technical field [0001] The invention belongs to the technical field of analysis and detection of nanomaterials for labeling biological systems, and more specifically relates to a method for detecting the interaction effect of heavy metal ions such as mercury and copper with cells by using the fluorescence intensity changes of cells after quantum dots mark cells with fluorescence. It can be used for in situ real-time detection of the content of heavy metal ions such as mercury and copper in cells, and it can also be used for research on the influence of heavy metal ions such as mercury and copper on the physiological activities of cells and the research on the transport pathways of heavy metal ions such as mercury and copper by cells. Background technique [0002] Cells are the basic structural units of living organisms, and various physiological functions and reactions of living things are based on cells and their products. Most heavy metal ions exist in the body at trace or...

Claims

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Application Information

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