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Scanning Electron Microscopy

Abstract

Bullfrog heart muscle trabecula are shock-frozen in liquid propane cooled by liquid nitrogen and then processed for X-ray microanalysis in two different ways: 1. Freeze-drying followed by vacuum embedding. 2. Cryoultramicrotomy and freeze-drying.

Stained sections of freeze-dried, embedded tissue exhibit detailed ultrastructure, but are useless for X-ray microanalysis. Unstained, dry cut plastic-sections are suitable for X-ray microanalysis, but the ultrastructure appears faint. Higher electron optical contrast and peak-to-background ratio of X-ray spectra are generally obtained in freeze-dried cryosections. Both preparation methods show that the X-ray spectra are influenced by the quality of cryofixation. The phosphorus/potassium ratio in nuclei increases with increasing ice crystal size.

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