AAT Bioquest

How does calcein AM work?

Posted July 22, 2020


Calcein AM is a non-fluorescent cell-permeable derivate of calcein that is widely used in cell viability measurement. The carboxylic acid groups on calcein are modified with AM (acetomethoxy) groups, which endows calcein AM with high hydrophobicity, facilizing its penetration through cell membrane. Once inside cell, AM groups are hydrolyzed by intracellular esterases. The fluorescent calcein molecule is restored, which is trapped in the cell and emits strong green fluorescence. Since dead cells lack esterase activity, only live cells are labeled and detected. The fluorescence intensity is proportional to esterase activity, thereby this assay is used to evaluate the cell viability.

Additional resources

Calcein, AM *UltraPure grade* *CAS 148504-34-1*

Live or Dead™ Cell Viability Assay Kit *Green/Red Dual Fluorescence*

Cell Meter™ Cell Viability Assay Kit *Blue Fluorescence*

Riss, T. L., Moravec, R. A., Niles, A. L., Duellman, S., Benink, H. A., Worzella, T. J., & Minor, L. (2016). Cell viability assays. In Assay Guidance Manual [Internet]. Eli Lilly & Company and the National Center for Advancing Translational Sciences.

Stoddart, M. J. (2011). Cell viability assays: introduction. In Mammalian cell viability (pp. 1-6). Humana Press.

Calcein AM Viability Dyes and Assay Kits

Cell Viability Assays