Van de Kant H J, De Rooij D G, Boon M E
Department of Cell Biology, State University of Utrecht, Medical School, The Netherlands.
Histochem J. 1990 Jun-Jul;22(6-7):335-40. doi: 10.1007/BF01003166.
A morphological and morphometrical study was performed on testicular cells after microwave stabilization of the tissue while immersed in phosphate buffered saline (PBS), 0.9 NaCl or Tris-HCl. Fixation in Carnoy's fluid without irradiation was chosen as a control chemical fixation method. After microwave stabilization or chemical fixation, the testes were embedded in paraffin or in plastic (glycolmethacrylate). An excellent morphology, comparable to that after chemical fixation in Carnoy's fluid, was observed in the plastic sections of tissue irradiated in PBS or NaCl, even when the sections were subsequently treated with an aggressive reagent at high temperature, required for the Feulgen reaction. The nuclear area of the microwave-stabilized Sertoli cells was 37-46% smaller in haematoxylin-eosin stained, paraffin sections in comparison with that in the glycolmethacrylate sections. The microwave-stabilized, paraffin-embedded tissue was much more vulnerable to the hot HCl treatment of the Feulgen staining than the chemically fixed tissue, resulting in an additional 10-20% decrease in nuclear size. The latter finding is particularly important for quantitative microscopy, where the Feulgen staining method is often employed.
对浸于磷酸盐缓冲盐水(PBS)、0.9%氯化钠或三羟甲基氨基甲烷盐酸盐(Tris-HCl)中的组织进行微波固定后,对睾丸细胞进行了形态学和形态计量学研究。选择未照射的卡诺氏液固定作为对照化学固定方法。微波固定或化学固定后,将睾丸包埋于石蜡或塑料(乙二醇甲基丙烯酸酯)中。在PBS或氯化钠中照射的组织的塑料切片中,观察到了与在卡诺氏液中化学固定后的形态相当的良好形态,即使这些切片随后在高温下用费尔根反应所需的侵蚀性试剂处理。苏木精-伊红染色的石蜡切片中,微波固定的支持细胞的核面积比乙二醇甲基丙烯酸酯切片中的小37%-46%。与化学固定的组织相比,微波固定、石蜡包埋的组织在费尔根染色的热盐酸处理中更易受损,导致核大小额外减小10%-20%。后一发现对于经常采用费尔根染色法的定量显微镜检查尤为重要。