Garland C D, Lee A, Dickson M R
J Microsc. 1979 Jul;116(2):227-42. doi: 10.1111/j.1365-2818.1979.tb00204.x.
Scanning electron microscopy (SEM) has become a popular means of studying micro-organisms which associate with surfaces. However, as yet no detailed examination has been made of the influence of specimen preparation on the number of organisms finally seen on the SEM screen. In this investigation critical assessment is made of the influence of a wide range or preparative factors on the preservation of filamentous bacteria associated with the epithelial surfaces of rat intestine. Organisms were quantitated using a rigorous counting method (transect line analysis); statistical testing of these counts enabled the comparison of different preparative factors. The composition of the fixative was found to significantly influence the number of organisms preserved; of the fifteen fixatives studied, Karnovsky's fixative with ruthenium red best preserved surface-associated organisms. The influence of other factors on the number and appearance of preserved organisms was also examined. These factors included the washing of specimens prior to fixation, the storage of fixed specimens, and the handling and storage of critical point dried specimens. The results are discussed with reference to the optimal methods for preparing specimens for SEM.
扫描电子显微镜（SEM）已成为研究与表面相关的微生物的一种常用手段。然而，目前尚未对标本制备对最终在扫描电子显微镜屏幕上看到的生物体数量的影响进行详细研究。在这项研究中，对一系列广泛的制备因素对与大鼠肠道上皮表面相关的丝状细菌保存的影响进行了严格评估。使用严格的计数方法（横断面线分析）对生物体进行定量；对这些计数进行统计测试能够比较不同的制备因素。发现固定剂的组成对保存的生物体数量有显著影响；在所研究的15种固定剂中，含钌红的卡诺夫斯基固定剂能最好地保存与表面相关的生物体。还研究了其他因素对保存的生物体数量和外观的影响。这些因素包括固定前标本的洗涤、固定标本的储存以及临界点干燥标本的处理和储存。结合用于扫描电子显微镜标本制备的最佳方法对结果进行了讨论。
