In Situ Electron-Microprobe Analyses of Tissue Cultures Grown on Glass

Abstract

The electron microprobe has found several applications in medicine and biology. These were recently reviewed by Tousirnis. He and others have shown it to be useful for the localization of Ca, P, Fe, Ag, Zn, Cr, and Sn in normal and diseased tissues. The majority of these studies were conducted on histological sections, several cells thick, with the cells cut in different planes, and overlapping one another.This paper reports on the analysis of cells grown in a horizontal monolayer. The cells are left intact, and are analyzed on the coverslip on which they are grown. L-929 fibroblasts and KB carcinoma cells were grown on 10 x 50-mm Bellco glass coverslips in Leighton tubes. The culture medium was Hanks' Basal medium of Eagle, supplemented with penicillin, streptomycin, and 10 percent calf serum. The cultures were incubated at 37 C in a horizontal position for periods up to 3 days. They were then fixed with absolute methanol for 10 seconds, washed with several changes of distilled water, and dried. The cultures were prepared for analysis by vacuum deposition of 100 A of elemental carbon.