Name: Overview of Cell Viability Assays
Creator: AAT Bioquest
License: https://www.aatbio.com/tou.html

Overview of Cell Viability Assays

Parameter	Assay/Reagents	Mode	Instrument	Principle
Membrane Integrity	Trypan Blue vital stains	Colorimetric	Hemocytometer	Trypan Blue, Trypan UltraBlue™, Trypan Red Plus™, and Trypan Purple™ are membrane-impermeant vital stains. Live cells with uncompromised membrane integrity exclude these dyes and remain unstained, whereas dead cells do not. The percentage of viable cells equals the number of unstained cells divided by the total number of cells (stained and unstained) multiplied by 100.
Membrane Integrity	Membrane-Impermeant DNA dyes	Fluorescence	FC, FM, or MR	Cell-permeable DNA dyes, such as propidium iodide, 7-AAD, or Nuclear DCS1™ dyes, enter dead cells with compromised membranes, whereby they bind to DNA and fluoresce brightly. Viable cells exclude these dyes and remain unstained.
Membrane Integrity	Cell Meter™ Fixable Viability dyes	Fluorescence	FC or FM	Cell-impermeable, amine-reactive dyes: in live cells, Cell Meter™ Fixable Viability dyes bind to primary amines on cell surface proteins amine and fluoresce dimly. In dead cells with damaged membranes, these dyes enter the cytoplasm and bind to many intracellular amine groups, resulting in significant fluorescence. The fluorescence intensity difference allows you to discriminate between live and dead cells.
Enzymatic activity	Calcein AM dyes	Fluorescence	FC, FM, or MR	Cell-permeable fluorogenic esterase substrates for identifying live cells. These substrates diffuse freely and noninvasively across the plasma membrane of live cells. Once inside, cytosolic esterases in active cells rapidly hydrolyze the non-fluorescent calcein AM substrates into brightly fluorescent calcein products retained by live cells with intact plasma membranes. The fluorescence intensity is proportional to the number of viable cells.
Metabolic activity	Tetrazolium dyes (WST and MTT)	Colorimetric	MR	Chromogenic substrates measure cellular reduction potential in metabolically active cells to evaluate viability. Active dehydrogenases and reductases in live, healthy cells reduce tetrazolium dyes from a colorless complex to a brightly purple-colored formazan product. Absorbance is measured at OD = 570 nm to determine viability.
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This online resource may be cited as follows

MLA	"Quest Database™ Overview of Cell Viability Assays." AAT Bioquest, Inc., 6 Dec. 2025, https://www.aatbio.com/data-sets/overview-of-cell-viability-assays.
APA	AAT Bioquest, Inc. (2025, December 6). Quest Database™ Overview of Cell Viability Assays. AAT Bioquest. https://www.aatbio.com/data-sets/overview-of-cell-viability-assays.
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