Institute of Neurobiology, Biomedical Research Center of the Slovak Academy of Sciences, Šoltésovej 4, 040 01 Košice, Slovakia.
Institute of Experimental Physics, Slovak Academy of Sciences, Watsonova 47, 040 01 Košice, Slovakia.
Neurosci Lett. 2021 Apr 17;750:135767. doi: 10.1016/j.neulet.2021.135767. Epub 2021 Feb 23.
We present a method that allows preparing histological sections from large blocks of nervous tissue embedded in epoxy resin. Resin-embedding provides excellent resolution especially for the myelin-rich white matter and is often being used for visualizing the myelinated axons in peripheral nerves. However, because of the limited penetration of the reagents, only very small tissue specimens can be processed in this way. Here, we describe a method that enables to embed large specimens and their sectioning on a standard sliding microtome. To process the large specimens, modifications in several steps of the processing technique had to be made. In this paper we demonstrate, that with this technique 1-3 μm thick transversal sections can be prepared from the resin-embedded specimens as large as rat brain hemisphere. Such a large section allows simultaneously: 1.) overviewing and delineating the gross anatomical structures, and 2.) observing the subcellular details at the highest possible optical magnifications. Such a large section with excellent resolution allows application of unbiased stereological methods and reliable quantification of very small objects within the area of interest.
我们提出了一种方法，可用于从包埋在环氧树脂中的大块神经组织中制备组织切片。树脂包埋可提供出色的分辨率，尤其适用于富含髓磷脂的白质，常用于可视化周围神经中的髓鞘化轴突。但是，由于试剂的渗透有限，只能以这种方式处理非常小的组织标本。在这里，我们描述了一种能够嵌入大标本并在标准滑动切片机上进行切片的方法。为了处理大标本，必须对处理技术的几个步骤进行修改。在本文中，我们证明，使用该技术可以从大鼠脑半球大小的树脂包埋标本中制备 1-3μm 厚的横向切片。这样一个大的切片可以同时：1.）概述和描绘大体解剖结构，和 2.）以尽可能高的光学放大倍数观察亚细胞细节。这种具有出色分辨率的大切片可用于应用无偏立体学方法和对感兴趣区域内非常小的物体进行可靠定量。
