MTT assay | Cell Viability and Cytotoxicity determination using MTT assay
Understanding MTTSA: A Colorimetric Assay for Cell Viability
Introduction to MTTSA
- The video introduces MTTSA, a colorimetric assay used to assess cell viability based on the reduction of tetrazolium dye by NADPH dependent cellular oxidoreductase enzymes.
- MTT (yellowish tetrazolium salt) is reduced to formazan (violet), indicating viable cells; non-viable cells do not undergo this reaction.
Applications of MTTSA
- MTTSA is primarily used for assessing cell viability and measuring cytotoxicity levels after treatment with therapeutic agents.
- It can also evaluate cytostatic activity, determining whether there is increased proliferation in cells treated with toxic materials.
Performing the MTTSA
- Cells must be plated at least 24 hours prior to conducting the assay. The MTT reagent, which is light-sensitive, should be dissolved in PBS and added in a dark setting.
- After incubating the plate with MTT for 4 hours, DMSO is added and incubated again for 2 hours before monitoring absorbance at 519 nm.
Data Interpretation
- A standard curve is generated using known cell numbers against their absorbance values, allowing extrapolation of unknown samples' cell numbers.
- An example involving cancer cell lines illustrates how treatment with a new compound can reduce cell viability, suggesting potential anti-cancer properties.
Summary of Key Learnings