Method for quantitatively measuring cell spreading rate based on idealized cell radius

A quantitative measurement and cell technology, applied in measurement devices, material analysis by optical means, instruments, etc., can solve the problems of difficult, rough, and no quantitative measurement of spreading rate of cells, and achieve the effect of simple calculation method and simple measurement equipment.

Inactive Publication Date: 2012-06-20
NANCHANG UNIV
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Problems solved by technology

However, this method of quantitatively measuring cell spreading is very rough: first, determining the three states of non-spreading, half-spreading, and fully spreading is very subjective, and there is no quantifiable standard. After all, each cell or even each individual Cells may have different shapes or adherent areas when they are fully spread. It is very difficult to judge whether a cell is fully spread, that is, there will be a certain deviation
Secondly, the semi-spreading state actually includes many intermediate states. Therefore, only dividing cell spreading into three states cannot quantitatively compare the changes of cell spreading over time.
Furthermore, quantifying cell spreading by counting cell number or percentage of number is an indirect method rather than a direct measure of spreading rate quantitatively

Method used

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  • Method for quantitatively measuring cell spreading rate based on idealized cell radius
  • Method for quantitatively measuring cell spreading rate based on idealized cell radius

Examples

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Effect test

Embodiment 1

[0016] Example 1: Quantitative measurement of cell spreading.

[0017] The suspended human umbilical vein endothelial cells were planted on a 24-well plate containing culture medium and placed in CO 2 Incubator (37 0 C, 5% CO 2 ) cultured. After 6 hours, 24 hours, and 48 hours of culture, observe and obtain cell morphology images with a confocal microscope, and measure the adherent area of ​​all individual cells in the image and the average adherent area of ​​all cells, through the formula provided above The average spreading rate of cells for different time periods was calculated. Finally, it was calculated that the average idealized radius at 6 hours, 24 hours and 48 hours during cell spreading was 12.95, 18.19 and 19.01 microns respectively; Over time, the average cell spreading rate (radius change per unit time) was 2.16, 0.29 and 0.03 μm / hour, respectively. The data showed that the cells spread the fastest within 0-6 hours, and the spreading speed gradually decreased...

Embodiment 2

[0018] Example 2: Quantitative measurement of drug effects on cell spreading.

[0019] The suspended human umbilical vein endothelial cells were planted on a 24-well plate containing culture medium and placed in CO 2 Incubator (37 0 C, 5% CO 2 ) cultured. After 24 hours of culture, use a confocal microscope to obtain cell morphology images, then add 20 ug / ml Ox-LDL (oxidized low-density lipoprotein) to the cell solution, and continue to culture for 48 hours, and at 6 hours and 12 hours respectively Topographic images of the cells were acquired using a confocal microscope at , 24, and 48 hours. And measure the adherent area of ​​all single cells in the image and the average adherent area of ​​all cells, and calculate the idealized radius and average spreading rate of cells in different time periods through the formula provided above. Finally, the effect of the drug on cell spreading was calculated: before adding the drug, the idealized radius of the cell was about 20.04 mic...

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Abstract

The invention relates to a method for quantitatively measuring cell spreading rate based on idealized cell radius. The method is characterized by comprising the steps of: planting adherent cells in a suspended state on culture holes or plates, capturing the morphology of the cells at different time points by using imaging equipment during the cell adhering and spreading process, measuring the adhering area of each cell, idealizing the cells before and after spreading to be round, and calculating the cell spreading rate (growth rate of the cell radius within unit time). According to the invention, the idealized cell radius is used as a standard, the spreading rate is calculated according to the cell spreading time and the change of the cell radius, and the defects of the traditional method are solved; and compared with the traditional cell spreading measuring method, the method has the advantages that the method is more accurate, the required measuring equipment is simple, and the calculation method is simple and feasible, and can be integrated to measuring equipment.

Description

technical field [0001] The invention belongs to the field of biotechnology, and is a method for measuring the spread rate of adherent cells and the influence of various drugs on the cell spread rate. technical background [0002] Cell spreading is one of the important physiological activities or functions of adherent cells, and many human physiological activities or diseases are related to cell spreading. For example, vascular endothelial cells build the inner wall of blood vessels or repair the damaged vascular endothelial layer through cell spreading. Therefore, changes in endothelial cell spreading are often related to the occurrence, development and tumor metastasis of cardiovascular diseases, and the effect of drugs on cell spreading It must also be one of the important indicators in in vitro cell tests to evaluate the efficacy of drugs. [0003] Unfortunately, the mechanism of cell spreading, especially the effect of drugs on cell spreading, is not well understood, an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/84
Inventor 陈勇邵文祥黄洁曾芳发
Owner NANCHANG UNIV
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