Abstract
A simple method for freeze-substitution is described. Substitution takes three days in a solution of osmium tetroxide in acetone and the material is subsequently embedded in araldite. The fixation obtained is comparable to that given by glutaraldehyde. The method is being used to study the ultrastructure of maize root tips, phloem and yeast. As fixation of the material only occurs after dehydration in acetone, the preservation of the various membrane systems of the cell is different from that seen in conventionally fixed cells. As the exact time of fixation by freezing is known, the method is also useful for the study of cells in which biochemical reactions occurring during conventional fixation result in ultrastructural changes.
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