Johnson E C, Hamer J D
J Reprod Fertil. 1975 Jan;42(1):95-104. doi: 10.1530/jrf.0.0420095.
Four methods of manipulating mouse uterine tissue during fixation, preparatory to examination of the luminal epithelium by scanning electron microscopy (S.E.M.), are described and the results assessed. The appearance of the epithelium varies according to the technique used, the choice of method depending on the type of information required from the S.E.M. study. Surface topography is preserved in a condition most closely resembling that of fresh tissue by opening the uterus and fixing the tissue flat. Critical examination of cellular and subcellular surface detail, however, depends on adequate spreading of the epithelial layer during fixation by distension of the uterine lumen under positive pressure. The method of tissue manipulation has a more profound effect on the appearance of the sample than any of the dehydration techniques which follow. For routine S.E.M. examinations at medium magnifications (smaller than X10,000), specimens which were air dried from alcohol, acetone or ether, or freeze dried, gave satisfactory results. Air drying from amyl acetate, and CO2 critical-point drying gave superior results at higher magnifications (greater than X10,000) with better preservation of individual microvilli.
本文描述了在通过扫描电子显微镜(S.E.M.)检查腔上皮之前,在固定过程中处理小鼠子宫组织的四种方法,并对结果进行了评估。上皮的外观因所使用的技术而异,方法的选择取决于S.E.M.研究所需的信息类型。通过打开子宫并将组织平铺固定,表面形貌得以保存在最接近新鲜组织的状态。然而,对细胞和亚细胞表面细节的严格检查取决于在正压下通过子宫腔扩张在固定过程中上皮层的充分铺展。组织处理方法对样品外观的影响比后续的任何脱水技术都更为深远。对于中等放大倍数(小于X10,000)的常规S.E.M.检查,从酒精、丙酮或乙醚中空气干燥或冷冻干燥的标本给出了满意的结果。从乙酸戊酯中空气干燥以及CO2临界点干燥在更高放大倍数(大于X10,000)下给出了更好的结果,且单个微绒毛的保存更好。