The PIT method: an automatedin vitrotechnique for drug toxicity testing

✍ Scribed by Rob Lambalgen; Peter Lelieveld

• Book ID: 104662279
• Publisher: Springer US
• Year: 1987
• Tongue: English
• Weight: 309 KB
• Volume: 5
• Category: Article
• ISSN: 0167-6997
• DOI: 10.1007/bf00203541

✦ Synopsis

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 IDs0 values, making use of adequate growth and no-growth controls.