Read N D, Jeffree C E
Institute of Cell and Molecular Biology, University of Edinburgh, U.K.
J Microsc. 1991 Jan;161(Pt 1):59-72. doi: 10.1111/j.1365-2818.1991.tb03073.x.
A review of low-temperature scanning electron microscopy (LTSEM) with regard to preparation protocols, specimen preservation, experimental approaches, and high-resolution studies, is provided. Preparative procedures are described and recent developments in methodologies highlighted. It is now well established that LTSEM, for most biological specimens, provides superior specimen preservation than does ambient-temperature SEM. This is because frozen-hydrated samples retain most or all of their water, are rapidly immobilized and stabilized by cryofixation, and are not exposed to chemical modification or solvent extraction. Nevertheless, artefacts in LTSEM are common and most arise because frozen-hydrated specimens contain water. LTSEM can be used as a powerful experimental tool. Advantages of employing LTSEM for this purpose and ways in which it can be used for novel experimentation are discussed. The most exciting development in recent years has been high-resolution LTSEM. The advantages, problems and requirements for this approach are defined.
本文对低温扫描电子显微镜（LTSEM）在制备方案、样本保存、实验方法及高分辨率研究方面进行了综述。文中描述了制备程序，并强调了方法学的最新进展。现已充分证实，对于大多数生物样本而言，LTSEM比常温扫描电子显微镜能提供更优质的样本保存效果。这是因为冷冻水合样本保留了大部分或全部水分，通过冷冻固定迅速固定并稳定下来，且未受到化学修饰或溶剂萃取的影响。然而，LTSEM中的伪像很常见，且大多是由于冷冻水合样本含有水分所致。LTSEM可作为一种强大的实验工具。文中讨论了为此目的使用LTSEM的优势以及它可用于新实验的方式。近年来最令人兴奋的进展是高分辨率LTSEM。文中定义了这种方法的优势、问题及要求。
