Morgenstern E
Fachbereich Theoretische Medizin, Universität des Saarlandes, Homburg, Saar.
Acta Histochem Suppl. 1977;Suppl 18:259-64.
Results obtained by conventional techniques and freeze-ultramicrotomie have been compared. Ultrathin frozen sections of platelets, which are fixed by glutaraldehyde, freeze-protected by polyvinyl-pyrrolidone and encapsulated in gelatin show a well preserved fine structure after staining with buffered OsO4, phosphotungstic, phosphomolybdic or ammoniummolybdic acid. The preservation of polysaccharides or mucopolysaccharides depends on a suitable fixation, that of glycogen in certain cases depends on a double fixation (glutaraldehyde and OsO4). Cytoplasmic nucleoproteins like ribosomes are visible only after drying of sections and staining with uranyl-acetate. Lipids, both membrane lipoproteins as well as lipids in the matrices of cell organelles, are well preserved. For example the zinc-iodide osmiumtetroxide reaction yields positive results in frozen sections contrary to such obtained by conventional methods. Furthermore, the extraction of altered lipids after staining with dyes like acridine orange does not take place with this method. For this reasons the freeze-ultramicrotomy seems to be an useful alternative method.
已对通过传统技术和冷冻超薄切片术获得的结果进行了比较。用戊二醛固定、聚乙烯吡咯烷酮进行冷冻保护并包埋在明胶中的血小板超薄冷冻切片,在用缓冲的四氧化锇、磷钨酸、磷钼酸或钼酸铵染色后,显示出保存良好的精细结构。多糖或粘多糖的保存取决于合适的固定,某些情况下糖原的保存取决于双重固定(戊二醛和四氧化锇)。细胞质核蛋白如核糖体仅在切片干燥并用醋酸铀染色后可见。脂质,包括膜脂蛋白以及细胞器基质中的脂质,都保存得很好。例如,碘化锌四氧化锇反应在冷冻切片中产生阳性结果,这与传统方法获得的结果相反。此外,用吖啶橙等染料染色后,这种方法不会发生改变的脂质的提取。基于这些原因,冷冻超薄切片术似乎是一种有用的替代方法。