Electron beam-induced changes in vitreous sections of biological samples

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Abstract

Summary

Nonpretreated high pressure frozen samples of Zea mays, cartilage and human erythrocytes were cryosectioned and observed at 110 K in a cryoelectron microscope. Changes induced by medium doses of electron irradiation (< 10 ke nm−2) are described. After some ke nm−2, the most conspicuous cutting artefacts are erased to a large extent and the visibility of the cell organelles is improved. The sections, compressed in the cutting direction by the sectioning process, shrink once more, in the same direction, when irradiated. This shrinkage depends on the section support and on how the section is adsorbed to it. Shrinkage is not uniform; it is most pronounced in mitochondria, condensed chromatin and nucleolus. This differential shrinkage improves the visibility of major structures on the section and, as a result, 'nicer' images are recorded. However, this apparent improvement is a beam-induced artefact that must be paired with a loss of high resolution information.

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