The electron probe x-ray microanalysis of the adult mammalian cardiac muscle.

Abstract

The main objective of this project was to master the electron probe x-ray microanalysis technique to permit one to obtain physiologically meaningful quantitative elemental profiles (Na, Cl, K, P, Mg, Ca & S) for the components of a given cell. The techniques which had to be mastered were: preparation of mammalian (rat) cardiac muscle; 'rapid' cryofixation using a Reichert-Jung MM80E 'impact freezer'; cryosectioning using a Reichert-Jung FC4 cryoultramicrotome; transfer and freeze-drying; electron probe x-rays collection using the EDAX 9100 Series Energy Dispersive X-ray Analysis Systems attached to a Philips TEM 420; quantitative analysis using the Hall equation and thin membrane aminoplastic standards. Different methods of transfer and freeze-drying were compared. Method I-a. Cryotransfer to the electron microscope using a modified Philips Cryotransfer system, freeze-drying in the column of the electron microscope, x-rays collection at low temperature; Method I-b. Cryotransfer to the electron microscope using a modified Philips Cryotransfer system, freeze-drying in the column of the electron microscope, x-rays collection at ambient temperature; Method II. Cryotransfer to a vacuum chamber (Edward-Coating System E306 A) using a precooled metal carrier, freeze-drying in a vacuum chamber while warming at ambient temperature, transfer and x-rays collection at ambient temperature. (Abstract shortened by UMI.)