MTT What is MTT and what is MTT method MTT, trade name thiazole blue Full name is 3-(4,5)-dimethylthiahiazo(z-y1)-3,5-di-phenytetrazoliumromide Chinese name is 3-(4,5-dimethylthiazole-2)-2,5-diphenyl Tetrazolium bromide, a yellow color dye, is well known for its application to the detection of cell viability. In cell culture, MTT is more than just a chemical, it is an experimental method. MTT principle MTT colorimetry is a method for detecting cell survival and growth. The detection principle is that succinate dehydrogenase in living cell mitochondria can reduce exogenous MTT to water-insoluble blue-purple crystalline formazan (Formazan) and deposit in cells, while dead cells do not have this function. Dimethyl sulfoxide (DMSO) can dissolve the hyperthyroidism in the cells, and its light absorption value is measured by a microplate reader at a wavelength of 490 nm, which can indirectly reflect the number of living cells. The amount of MTT crystal formation is proportional to the number of cells in a certain number of cells. The method has been widely used for the detection of activity of some biologically active factors, large-scale anti-tumor drug screening, cytotoxicity test, and tumor radiosensitivity measurement. It is characterized by high sensitivity and economy. MTT use MTT has two main uses 1. Determination of cytotoxicity of drugs in vitro (including other treatments such as radiation); 2. Cell proliferation and cell viability assay. The detection principle is as described above, and the number of living cells is determined based on the measured absorbance value (OD value). The larger the OD value, the stronger the cell activity (if the drug toxicity is measured, the drug toxicity is smaller). MTT experimental steps Ordinary MTT method experimental steps: 1: Inoculation of cells: A single cell suspension was prepared by using a culture solution containing 10% fetal calf serum, and inoculated into a 96-well plate at a volume of 1000-10000 cells per well, each volume of 200 ul. 2 : Cultured cells: cultured for 3-5 days under normal culture conditions (the culture time can be determined according to the purpose and requirements of the test). 3: Coloration: After 3-5 days of culture, add MTT solution (5 mg/ml in PBS, pH=7.4) 20 ul per well. Continue to incubate for 4 h. The culture was terminated, and the culture supernatant in the well was carefully discarded. After the suspension was centrifuged, the culture supernatant in the well was discarded. 150 ul of DMSO was added to each well and shaken for 10 min to allow the crystals to fully melt. 4: Colorimetric: Select the wavelength of 490 nm, measure the light absorption value of each well on the microplate reader, record the result, plot the cell growth curve with time as the abscissa and absorbance as the vertical coordinate. MTT detection method The results of MTT experiments were commonly read by microplate readers. The cell growth status was judged by comparing the OD values ​​of different wells, which was used as an important basis for comparing drug toxicity or cell activity. Punch Lithoclast Set,Energetically Lithotrite Endoscope,Urology Endoscopic Surgical Instruments,Urological Sigmoidoscope Set ZHEJIANG SHENDASIAO MEDICAL INSTRUMENT CO.,LTD. , https://www.sdsmedtools.com