Cell Viability Assays

TLDR

The chapter provides an introductory overview of common assays for estimating viable cell numbers in multi‑well plates. It describes plate‑reader based assays—including colorimetric tetrazolium, resazurin reduction, protease‑based fluorescence, luminogenic ATP, and a novel real‑time live‑cell monitoring assay—that measure marker activity linked to viable cell number. These assays enable measurement of cell proliferation, assessment of compound cytotoxicity, and multiplexing as internal controls in other cell‑based assays.

Abstract

This chapter is an introductory overview of the most commonly used assay methods to estimate the number of viable cells in multi-well plates. This chapter describes assays where data are recorded using a plate-reader; it does not cover assay methods designed for flow cytometry or high content imaging. The assay methods covered include the use of different classes of colorimetric tetrazolium reagents, resazurin reduction and protease substrates generating a fluorescent signal, the luminogenic ATP assay, and a novel real-time assay to monitor live cells for days in culture. The assays described are based on measurement of a marker activity associated with viable cell number. These assays are used for measuring the results of cell proliferation, testing for cytotoxic effects of compounds, and for multiplexing as an internal control to determine viable cell number during other cell-based assays.