A method for measuring cytoplasm viscosity based on quantum dot three-dimensional tracing

A quantum dot, cytoplasmic technology, applied in measurement devices, indirect flow characteristic measurement, flow characteristics, etc., can solve the problems of inappropriate real-time tracking of quantum dots, inability to intuitively understand the three-dimensional motion characteristics of molecules, etc., and achieve high real-time performance and positioning. The effect of precision, accurate cytoplasmic viscosity

Active Publication Date: 2018-02-16
XI AN JIAOTONG UNIV
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Problems solved by technology

At present, scholars have used it as a probe of intracellular viscosity to reflect the value of cell viscosity through fluorescence intensity changes, fluorescence anisotropy and fluorescence lifetime, but this method cannot intuitively understand the three-dimensional motion characteristics of molecules under specific cytoplasmic viscosity
In order to obtain the three-dimensional motion characteristics of molecules in the cytop...

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  • A method for measuring cytoplasm viscosity based on quantum dot three-dimensional tracing
  • A method for measuring cytoplasm viscosity based on quantum dot three-dimensional tracing
  • A method for measuring cytoplasm viscosity based on quantum dot three-dimensional tracing

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Embodiment Construction

[0031] The present invention will be described in further detail below in conjunction with the accompanying drawings.

[0032] A method for obtaining cytoplasmic viscosity based on three-dimensional tracking of quantum dots, comprising the following steps:

[0033] (1) Quantum dots label cells

[0034] This method takes HepG2 cells labeled with CdTe quantum dots as an example, adopts the liposome Lipo-2000 transfection method, and obtains the following operation process after multiple experimental parameter adjustments and optimizations:

[0035] (1) When the fourth-generation HepG2 cells are 80-90% confluent, digest them with 2.5% trypsin-EDTA, resuspend the culture medium, and count the cells. According to the counting results, dilute the cell density to 1.0× 10 5 cells / mL, take 1 mL of the cell suspension and inoculate it on a confocal culture dish (NEST) with a diameter of 20 mm. After culturing for 10 h, the cells are completely adhered to the wall; DMEM medium, CO 2 ...

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Abstract

A method for measuring cytoplasm viscosity based on quantum dot three-dimensional tracing is disclosed. The method includes firstly introducing quantum dots into cells through a lipid Lipo-2000 transfection manner to achieve cell labeling; subjecting the single and recognizable quantum dots dispersed in the cells to three dimensional positioning and tracing by adopting an optical modulating mannerthrough a build quantum dot three-dimensional imaging system; further solving mean square displacement values MSDs which are motion parameters of the quantum dots by utilizing three-dimensional motion information; selecting quantum dots meeting free diffusion characteristics according to mathematical characteristics of the MSDs; finally solving diffusion coefficient D of the quantum dots in cytoplasm according to the MSD values for all the quantum dots meeting free diffusion characteristics; and further acquiring the cytoplasm viscosity according to a Stokes-Einstein equation. In a state thatcells survive, the cytoplasm viscosity can be accurately acquired by the method, visual three-dimensional motion information of molecules in cytoplasm can be obtained, and technical bases for researching intracellular material transportation, signal transmission, metabolism, differentiation, and the like are provided.

Description

technical field [0001] The invention belongs to the technical field of micro-nano three-dimensional imaging, and in particular relates to a method for measuring cytoplasmic viscosity based on quantum dot three-dimensional tracing. Background technique [0002] Cytoplasm is the main site of cell life activities, and is the general term for all translucent, gelatinous, and granular substances surrounded by the cytoplasmic membrane except the nuclear region. The viscosity of the cytoplasm is the main factor affecting the diffusion of biomolecules. By obtaining the viscosity value of the cytoplasm, we can further understand important processes such as material transport, signal transmission, metabolism and differentiation in living cells. The current method for determining the viscosity of cytoplasm is to use a viscometer to measure the viscosity of cytoplasm by extracting the cytoplasm. However, after the cytoplasm is extracted from the cell body, its characteristics in the ce...

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Application Information

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IPC IPC(8): G01N11/10G01N11/00G01N13/00
CPCG01N11/00G01N11/10G01N13/00G01N2011/008G01N2013/003
Inventor 李本强阎贝江新兵刘红忠
Owner XI AN JIAOTONG UNIV
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