Department of Histology (Tissue Engineering Group), Faculty of Medicine, University of Granada, Granada, Spain.
Instituto de Investigación Biosanitaria, Ibs.GRANADA, Granada, Spain.
Methods Mol Biol. 2023;2566:187-203. doi: 10.1007/978-1-0716-2675-7_15.
Histochemical and fluorescence-based techniques enable the specific identification of myelin by bright-field or fluorescence microscopy. In this chapter, we describe four histological methods for the evaluation of myelin on peripheral nerve tissue sections. The first method combines the Luxol fast blue (LFB) technique with a modified Picrosirius staining contrasted with Harris hematoxylin, called MCOLL. This method simultaneously stains myelin, collagen fibers, and cell nuclei, thus giving an integrated overview of the histology, collagen network, and myelin content of the tissue in paraffin-embedded or cryosectioned samples. Secondly, we describe the osmium tetroxide method, which provides a permanent positive reaction for myelin as well as other lipids present in the tissue. The third method is the immunofluorescence-based detection of myelin proteins that allows to combine information about their expression status with other proteins of interest. Finally, the FluoroMyelin™ stains enable a fast detection of the myelin content that can be easily implemented in immunofluorescence staining panels for cryosectioned tissues. Together, this chapter provides a variety of methods to accurately identify myelin in different experimental approaches.
组织化学和荧光技术能够通过明场或荧光显微镜对髓鞘进行特异性识别。在本章中，我们描述了四种用于评估周围神经组织切片中髓鞘的组织学方法。第一种方法将洛索夫快速蓝（LFB）技术与改良的苦味酸天狼星红染色相结合，称为 MCOLL。该方法同时对髓鞘、胶原纤维和细胞核进行染色，从而为石蜡包埋或冷冻切片样本的组织学、胶原网络和髓鞘含量提供综合概述。其次，我们描述了锇四氧化物法，该方法为组织中存在的髓鞘和其他脂质提供永久性阳性反应。第三种方法是基于免疫荧光的髓鞘蛋白检测，它允许将关于其表达状态的信息与其他感兴趣的蛋白质结合起来。最后，FluoroMyelin™ 染色能够快速检测髓鞘含量，可轻松应用于冷冻切片组织的免疫荧光染色组中。总之，本章提供了多种方法来准确识别不同实验方法中的髓鞘。
