Raimondo Stefania, Fornaro Michele, Di Scipio Federica, Ronchi Giulia, Giacobini-Robecchi Maria G, Geuna Stefano
Department of Clinical and Biological Sciences, San Luigi Gonzaga School of Medicine, University of Turin, Turin 10043, Italy.
Int Rev Neurobiol. 2009;87:81-103. doi: 10.1016/S0074-7742(09)87005-0.
This paper critically overviews the main procedures used for carrying out morphological analysis of peripheral nerve fibers in light, confocal, and electron microscopy. In particular, this paper emphasizes the importance of osmium tetroxide post-fixation as a useful procedure to be adopted independently from the embedding medium. In order to facilitate the use of any described techniques, all protocols are presented in full details. The pros and cons for each method are critically addressed and practical indications on the different imaging approaches are reported. Moreover, the basic rules of morpho-quantitative stereological analysis of nerve fibers are described addressing the important concepts of design-based sampling and the disector. Finally, a comparison of stereological analysis on myelinated nerve fibers between paraffin- and resin-embedded rat radial nerves is reported showing that different embedding procedures might influence the distribution of size parameters.
本文批判性地概述了在光学显微镜、共聚焦显微镜和电子显微镜下对周围神经纤维进行形态学分析所使用的主要程序。特别是，本文强调了四氧化锇后固定作为一种可独立于包埋介质采用的有用程序的重要性。为便于使用所描述的任何技术，所有方案都详细列出。对每种方法的优缺点进行了批判性讨论，并报告了不同成像方法的实际应用指南。此外，还描述了神经纤维形态定量立体分析的基本规则，阐述了基于设计的抽样和均等分割体这两个重要概念。最后，报告了对石蜡包埋和树脂包埋的大鼠桡神经有髓神经纤维进行立体分析的比较结果，表明不同的包埋程序可能会影响尺寸参数的分布。
