Development of propane jet freezing methods for cultured cardiac cells

Abstract

Physiologically viable and synchronously beating heart cells can be grown in varying geometric configurations for parallel biochemical, electrophysiological, ultrastructural and x-ray microanalytical studies. Our objective in these experiments was to develop a method by which heart cells grown to confluency either as a sheet, spherical aggregate or polystrand could be rapidly frozen prior to cryosectioning and electron probe x-ray microanalysis.A single-sided propane jet freezer, which has been shown to achieve excellent freezing of confluent cell sheets prior to freeze fracture, was constructed according to Pscheid et al.; a specimen holder was also designed and constructed which facilitated rapid transfer (<10 sec) of the various cultured cells from media to the jet freezer (FIG. 1a,b).