EVALUATION OF THE TRYPAN BLUE TECHNIQUE FOR DETERMINATION OF CELL VIABILITY

TLDR

The study aims to evaluate the accuracy of trypan blue staining as an index of viable cell proportion and recommends defining the specific system before acceptance. The authors compared trypan blue staining results with growth potential assays in vitro and in vivo, and examined how trauma, freezing, and dispersal agents affect cell viability. Eosin and acridine orange proved relatively toxic; trypan blue staining correlates with cultivation but only yields about 85 % of cultivability values under optimal handling, and becomes grossly inaccurate after traumatic treatment.

Abstract

A study has been made to determine the accuracy of the trypan blue staining technique as an index of the proportion of viable cells in a cell population. Eosin and acridine orange were also tested in preliminary studies but were found to be relatively toxic. Cell cultures in vitro and tumor cell populations tested in vivo are used for direct comparisons between viability values obtained by staining cell populations with trypan blue, and values obtained by testing the growth potential of the same cell populations under optical conditions of cultivation. Data are included relative to the effects of trauma, freezing, and of various dispersal agents upon cell viability. The results indicate correlation but not absolute agreement between the two methods (staining and cultivation). Trypan blue values are shown to be 85% (or better) of cultivability values under optimal conditions of handling, but they are grossly inaccurate and unpredictable following traumatic treatment of cell populations. Before trypan blue staining is accepted as an index of cell viability, it is recommended that the specific system under study first be carefully defined.