Automated fluorescent analysis for drug-induced cytotoxicity assays

K. Funa, N. Dawson, P. B. Jewett, H. Agren, J. C. Ruckdeschel, P. A. Bunn, A. F. Gazdar

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

The human tumor clonogenic assay has been reported to predict for sensitivity of human tumors to a variety of drugs. However, this assay requires large numbers of viable cells, is time-consuming, and takes at least 2 weeks before results are available. To circumvent these problems, Weisenthal developed a microscope-based dye exclusion assay. Because this method is also time-consuming and subject to observer error, we have developed an automated method of quantitating drug cytotoxicity using a flow cytometric cell sorter (FCM). After incubation of drug-exposed tumor cells, acetaldehyde-fixed duck red blood cells (DRBC) are added. Dead tumor cells and the fixed DRBC are stained by the fluorescent dye propidium iodide, which penetrates dead cell membranes. A two-parameter analysis (cell size as measured by narrow angle light scatter vs propidium iodide fluorescence) enables determination of the live tumor cell:DRBC ratio. There was a strong correlation between the FCM method and manual counting (r = 0.958 for cell lines, r = 0.831 for fresh leukemic cells, P < 0.0001 in both cases). We conclude that the automatized FCM method gives compatible results to the manual dye exclusion assay and increases efficiency.

Original languageEnglish (US)
Pages (from-to)1147-1151
Number of pages5
JournalCancer Treatment Reports
Volume70
Issue number10
StatePublished - Dec 30 1986

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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