Peters K R
J Microsc. 1980 Apr;118(4):429-41. doi: 10.1111/j.1365-2818.1980.tb00292.x.
An exact method of preparation of soft biological specimens for electron microscopic analysis of surface fine structures is described. It allows routine preparations of fragile specimens for SEM and TEM imaging modes. With this procedure physical preparation parameters such as mechanical loads on the specimen surface or changes of temperature are controlled. The wet specimens are premounted in cheap disposable BEEM-containers or glass boats and are constantly kept under liquid in a closed system. The exchange of preparation media is performed continuously and, if necessary, over gradients. For comparative investigations with different EM-modes, at each step of the procedure parts of the specimens may be removed for individual processing. Conventionally prepared critical-point dried specimens are compared to those processed by the exchange technique and preservation of surface fine structures is demonstrated. Shadow-casted clathrin cages and stereo-replicas of virus infected cell cultures are shown in TEM preparations. For SEM, coverslip cell cultures and isolated glomerulus basement membranes are prepared and an additional flat embedding for TEM ultrathin sections is demonstrated.
本文描述了一种用于电子显微镜分析表面精细结构的软生物标本精确制备方法。它可用于为扫描电子显微镜（SEM）和透射电子显微镜（TEM）成像模式对易碎标本进行常规制备。通过此程序，可控制物理制备参数，如标本表面的机械负荷或温度变化。湿标本预先安装在廉价的一次性BEEM容器或玻璃舟中，并在封闭系统中始终保持在液体下。制备介质的交换持续进行，必要时可通过梯度进行。对于不同电子显微镜模式的比较研究，在程序的每个步骤中，可取出部分标本进行单独处理。将传统制备的临界点干燥标本与采用交换技术处理的标本进行比较，并展示表面精细结构的保存情况。在透射电子显微镜制备中展示了阴影投射的网格蛋白笼和病毒感染细胞培养物的立体复制品。对于扫描电子显微镜，制备了盖玻片细胞培养物和分离的肾小球基底膜，并展示了用于透射电子显微镜超薄切片的额外扁平包埋。
