Weil Marie-Theres, Ruhwedel Torben, Möbius Wiebke, Simons Mikael
Department of Cellular Neuroscience, Max-Planck Institute for Experimental Medicine, Göttingen, Germany.
Department of Neurogenetics, Max-Planck Institute for Experimental Medicine, Göttingen, Germany.
Curr Protoc Neurosci. 2017 Jan 3;78:2.27.1-2.27.18. doi: 10.1002/cpns.22.
Intracerebral injections are an invasive method to bypass the blood brain barrier and are widely used to study molecular and cellular mechanisms of the central nervous system. The administered substances are injected directly at the site of interest, executing their effect locally. By combining injections in the rat brain with state-of-the-art electron microscopy, subtle changes in ultrastructure of the nervous tissue can be detected prior to overt damage or disease. The protocol presented here involves stereotactic injection into the corpus callosum of Lewis rats and the cryopreparation of freshly dissected tissue for electron microscopy. The localization of the injection site in tissue sections during the sample preparation for transmission electron microscopy is explained and possible artifacts of the method are indicated. With the help of this powerful combination of injections and electron microscopy, subtle effects of the applied substances on the biology of neural cells can be identified and monitored over time. © 2017 by John Wiley & Sons, Inc.
脑内注射是一种绕过血脑屏障的侵入性方法，被广泛用于研究中枢神经系统的分子和细胞机制。所施用的物质直接注射到感兴趣的部位，在局部发挥作用。通过将大鼠脑内注射与最先进的电子显微镜技术相结合，可以在明显的损伤或疾病出现之前检测到神经组织超微结构的细微变化。这里介绍的方案包括对Lewis大鼠胼胝体进行立体定向注射，以及对新鲜解剖的组织进行冷冻制备以用于电子显微镜观察。文中解释了在透射电子显微镜样品制备过程中注射部位在组织切片中的定位，并指出了该方法可能出现的假象。借助注射和电子显微镜这一强大的组合，可以识别并随时间监测所施用物质对神经细胞生物学的细微影响。© 2017约翰威立国际出版公司
