AAT Bioquest

Which viability test is better: MTT or trypan blue?

Posted May 9, 2020


Each test has its advantages and limitations.

Trypan blue is a dye exclusion assay in which dead cells will take up the negatively charged dye due to their damaged membranes, while live cells exclude it. This is a rapid and simple assay and is good for analyzing membrane integrity. However, with this assay, cells that are still alive but losing their function will not be stained; there also may be counting errors.

MTT is a colorimetric assay in which viable cells reduce MTT into a purple precipitate formazan, which is then solubilized before measuring absorbance. MTT has higher sensitivity but can also overestimate cell viability and cannot be used with compounds such as polyphenols, ascorbic acid, vitamin A, coenzyme A, and DTT, which can interfere with the reduction of MTT to formazan.

Additional resources

Aslantürk, Ö. S. (2017). In vitro cytotoxicity and cell viability assays: Principles. Advantages, and Disadvantages.

Wang, P., Henning, S. M., & Heber, D. (2010). Limitations of MTT and MTS-based assays for measurement of antiproliferative activity of green tea polyphenols. PloS one, 5(4).

Trypan Blue, sodium salt *10 mM aqueous solution*

Cell Meter™ Colorimetric MTT Cell Proliferation Kit

MTT Assay