Quantitative Fluorescent Microassay for Identification of Antiproliferative Compounds<xref ref-type="fn" rid="FN2">2</xref>

Abstract

A reproducible and convenient microassay was developed by us to measure the antiproliferative activity of biological materials and anticancer drugs. The assay involved the labeling of target cells after their incubation with anticancer agents with the fluorescent vital dye hydroethidine at a dose of 28 micrograms/ml for 30 minutes. Once internalized into live cells, hydroethidine is dehydrogenated to ethidium, which then intercalates into DNA. Hydroethidine labeled the cells uniformly. The cells were lysed with detergent, and the relative fluorescence of this dye was monitored in a microfluorimeter at the speed of 30 seconds per 96-well plate. Because this assay is accurate and reproducible, does not present problems of disposal of labeling agents, and is extremely rapid, it should prove very useful for the screening of many potential anticancer agents with antiproliferative activity.