The PIT method: an automated in vitro technique for drug toxicity testing

An automated in vitro technique for drug toxicity testing is described. Human tumor cells were cultured for 2 days in 96-well microtiter plates before the addition of serial dilutions of drugs. At day 5 the cultures were terminated by the addition of a solution containing propidium iodide, ink and triton X-100 (PIT). Triton X-100 lysed all cells, which were subsequently stained by the DNA specific fluorescing propidium iodide. The ink effectively quenched all background fluorescence. The plates were read on a computer controlled automated microscope with a photomultiplier. The results showed a linear relationship between cell number and fluorescence intensity. Reproducible dose-response curves were obtained for 6 drugs tested. A computer program calculated ID50 values, making use of adequate growth and no-growth controls.