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Application of cell culture plate in MTT detection

Thu Aug 26 16:43:51 CST 2021

The cell culture plate is a kind of biological tool. It is divided into flat bottom and round bottom according to the shape. The number of wells includes 6 wells, 12 wells, 24 wells, 48 ​​wells, 96 wells, 384 wells, etc. It is mainly used for cell culture and can also be used for to measure protein concentration, 96-well plates are commonly used for MTT detection.

MTT method, also known as MTT colorimetric method, is a method for detecting cell survival and growth. The detection principle is that the succinate dehydrogenase in the mitochondria of living cells can reduce the exogenous MTT to water-insoluble blue-purple crystal formazan (Formazan) and deposit it in the cells, while dead cells have no such function. Dimethyl sulfoxide (DMSO) can dissolve formazan in cells, and its light absorption value is measured at 570nm with an enzyme-linked immunoassay, which can indirectly reflect the number of living cells. Within a certain range of cell numbers, the amount of MTT crystal formation is proportional to the number of cells. The detection steps are as follows:

1. Culture the cells in a 96-well cell culture plate. After the cell density is appropriate, add 50ul MTT to each well and continue to culture for 4 hours;

2. Suck away the culture medium containing MTT and add 150ul of DMSO. Vibrate horizontally for 10 minutes to fully dissolve the crystals. For suspended cells, after centrifugation, suck away the medium, and then add DMSO to dissolve;

3. Measure the absorbance at 550nm on the enzyme-linked immunoassay instrument (set a reasonable control group and blank group);

4. Cell survival rate=absorbance value of experimental group/absorbance value of control group*100%.

It should be noted during the operation that MTT is carcinogenic. If possible, try to wear the kind of transparent film gloves. The prepared MTT needs to be sterile. MTT is very sensitive to bacteria. It does not matter if you add to the 96-well cell culture plate without protecting it from light. , After all, the time is short, or you can turn off the lighting on the console.

The FAI climbed 5.9 percent year-on-year in the first 11 months of 2018, quickening from the 5.7-percent growth in Jan-Oct, the National Bureau of Statistics (NBS) said Friday in an online statement.

The key indicator of investment, dubbed a major growth driver, hit the bottom in August and has since started to rebound steadily.

In the face of emerging economic challenges home and abroad, China has stepped up efforts to stabilize investment, in particular rolling out measures to motivate private investors and channel funds into infrastructure.

Friday's data showed private investment, accounting for more than 60 percent of the total FAI, expanded by a brisk 8.7 percent.

NBS spokesperson Mao Shengyong said funds into weak economic links registered rapid increases as investment in environmental protection and agriculture jumped 42 percent and 12.5 percent respectively, much faster than the average.

In breakdown, investment in high-tech and equipment manufacturing remained vigorous with 16.1-percent and 11.6-percent increases respectively in the first 11 months. Infrastructure investment gained 3.7 percent, staying flat. Investment in property development rose 9.7 percent, also unchanged.