Protocol

Viability Staining of Mammalian Cell Cultures

This protocol was adapted from "Growth and Manipulation of Cells in Culture," Chapter 2, in Cells (eds. Spector et al.). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 1998.

INTRODUCTION

Various manipulations of cells, including passaging, freezing, and dissociation from primary tissue, can result in cell death. To determine the number of surviving cells in a population, exclusion of the dye trypan blue can be used. Normal healthy cells are able to exclude the dye, but trypan blue will diffuse into cells in which membrane integrity has been lost. This dye-exclusion method provides a rough estimate of cell viability and often does not distinguish within a 10%-20% difference. Additionally, cells that exclude dye are not necessarily capable of attachment and prolonged survival or proliferation.