Journal of tissue culture methods

, Volume 9, Issue 1, pp 7–9

A simple quantitative procedure using monolayer cultures for cytotoxicity assays (HTD/NR-90)

  • Ellen Borenfreund
  • James A. Puerner

DOI: 10.1007/BF01666038

Cite this article as:
Borenfreund, E. & Puerner, J.A. Journal of Tissue Culture Methods (1985) 9: 7. doi:10.1007/BF01666038


A sensitive quantitative procedure for assaying viable cells in monolayer cultures is described. The two-component test involves (a) microscopic screening for morphological alterations after an experimental protocol for cytotoxicity studies and (b) quantitation of surviving cells by incubation with the supravital dye neutral red, followed by colorimetric analysis of the dye extracted from the lysosomes of the viable cells. Both assays, conveniently carried out within the same culture, have been standardized for use with 96-well microtiter plates and automatic reading with a Dynatech spectrophotometric microplate reader. The test can be adapted for use with conventional spectrophotometers.

Key words

cytotoxicitymorphologyneutral redspectrophotometric analysis

Copyright information

© Tissue Culture Association, Inc 1985

Authors and Affiliations

  • Ellen Borenfreund
    • 1
  • James A. Puerner
    • 1
  1. 1.Laboratory Animal Research CenterThe Rockefeller UniversityNew YorkUSA