Unique method of tissue culture preparation for EM

Abstract

A simplified technique of growing monolayer cells for electron microscopic examination has been developed. Our procedure has eliminated many difficult steps and therefore is easier than those reported by others.Regular Beem capsules for routine embedding for electron microscopy were used(Fig. 1). Prior to tissue inoculation capsules were washed with 5% HCl and gas sterilized. A 0.5 ml cell suspension of MCF-7 cells (10,000/ml) was placed in the capsules and cells settled in the pyramid portion (Fig. 2). Culture medium was alpha-MEM with 10% FCS. Capsules were incubated at 37°C for 3 days. They were then fixed in 70% ethanol for 20 minutes and stained with crystal violet. The pyramid portion of the capsule was cut off and monolayer cell growth was confirmed by examination under a microscope (Fig. 3).