Morphometry of cell organelles on thick sections by high voltage electron microscopy

Abstract

Although the analysis of size distributions of cell organelles on the ultrathin sections is usually carried out by conventional electron microscopy, the procedure brings about some errors. The purpose of this paper is to describe a more useful method for morphometry by high voltage electron microscopy and to demonstrate how this method can be applied to practical problems.The pituitary,the kidney and the liver of a mouse and rats were used as materials. The pituitary gland was obtained from a male mouse. It was doubly fixed in 2.5% glutaraldehyde and 1% osmium tetroxide,dehydrated with a graded series of ethanol and embedded in Epon. The kidney and the liver were obtained from male rats treated with DEHP ( di-2-ethylhexyl phthalate),a peroxisomes proliferating agent. They were prefixed in 2.5% glutaraldehyde for 3 hours and were incubated in a DAB.pH 9.0,medium containing hydrogen peroxide for 60 minutes at 37°C. They were postfixed in 1% osmium tetroxide, dehydrated and embedded in Epon. Ultrathin (0.lμm), semi thin (0.2μm) and thick(0.5μm, 1.0μm) sections from the same Epon block were cut with an LKB Ultrotome respectively.