Klein H P, Stockem W
Microsc Acta. 1976 Nov;78(5):388-406.
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.
通过对两种含水量不同的变形虫(大变形虫、简单帆变形虫)进行研究,测试了通常用于扫描电子显微镜生物材料制备的三种方法。空气干燥导致细胞收缩和细胞变形。当从蒸气压逐渐增加的培养基中干燥标本时,可以获得更令人满意的表面结构保存效果。效力顺序为:乙醇、氯仿、异戊烷、乙醚、氟利昂11和氟利昂13。短干燥期被证明比长干燥期更有利。临界点干燥在两种变形虫物种中都提供了更好的细胞表面形态细节。然而,一些人为变化仍然可以检测到,如粘液层的小破裂和破坏。它们必须归因于固定和脱水过程。冷冻干燥被证明优于空气干燥和临界点干燥。通过这种方法制备的标本与活细胞相比,在细胞表面形态上没有可见差异。由于含水量相对较高,大变形虫的制备比简单帆变形虫更困难。