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  • Title: [Preparation of biological specimens for the scanning electron microscope (author's transl)].
    Author: Klein HP, Stockem W.
    Journal: Microsc Acta; 1976 Nov; 78(5):388-406. PubMed ID: 794634.
    Abstract:
    Three methods usually applied in preparing biological material for the scanning electron microscope were tested by the investigation of two species of amoebae with different content of water (Amoeba proteus, Vannella simplex). Air drying resulted in both the production of cell shrinkage and cell distortion. When the specimens were dryed from media with increasing vapour-pressure, more satisfactory preservation of surface structures could be obtained. The sequence of potency was: Ethanol, chloroform, isopentane, ethyl ether, freon 11 and freon 13. Short drying periods proved to be more favourable than long ones. Critical-point drying provided much better details of cell surface morphology in both amoebae species. However, some arteficial changes were still detectable as small breaks and destruction of the mucous layer. They must be attributed to the fixation and dehydration procedure. Freeze drying turned out to be superior to both air drying and critical-point drying. Specimens prepared by this method showed no visible differences in cell surface morphology compared to living cells. As a consequence of the relatively high content of water the preparation of A. proteus was more difficult than that of V. simplex.
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