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适应福尔马林固定人脑和光学显微镜的“单节”高尔基法。

The "single-section" Golgi method adapted for formalin-fixed human brain and light microscopy.

机构信息

Program in Neuroscience, Institute of Basic Sciences, Federal University of Rio Grande do Sul, R. Sarmento Leite 500, Porto Alegre RS 90050-110, Brazil.

出版信息

J Neurosci Methods. 2010 May 30;189(1):51-5. doi: 10.1016/j.jneumeth.2010.03.018. Epub 2010 Mar 27.

Abstract

The Golgi method has been used for over a century to describe the general morphology of neurons in the nervous system of different species. The "single-section" Golgi method of Gabbott and Somogyi (1984) and the modifications made by Izzo et al. (1987) are able to produce consistent results. Here, we describe procedures to show cortical and subcortical neurons of human brains immersed in formalin for months or even years. The tissue was sliced with a vibratome, post-fixed in a combination of paraformaldehyde and picric acid in phosphate buffer, followed by osmium tetroxide and potassium dicromate, "sandwiched" between cover slips, and immersed in silver nitrate. The whole procedure takes between 5 and 11 days to achieve good results. The Golgi method has its characteristic pitfalls but, with this procedure, neurons and glia appear well-impregnated, allowing qualitative and quantitative studies under light microscopy. This contribution adds to the basic techniques for the study of human nervous tissue with the same advantages described for the "single-section" Golgi method in other species; it is easy and fast, requires minimal equipment, and provides consistent results.

摘要

高尔基法已经使用了一个多世纪,用于描述不同物种神经系统中神经元的一般形态。加博特和索莫吉(1984 年)的“单切片”高尔基法和伊佐等人(1987 年)的修改能够产生一致的结果。在这里,我们描述了将福尔马林浸泡数月甚至数年的人脑皮质和皮质下神经元的程序。组织用振动切片机切片,然后在磷酸盐缓冲液中用多聚甲醛和苦味酸的混合物后固定,再用四氧化锇和重铬酸钾处理,“夹”在盖玻片之间,然后浸泡在硝酸银中。整个过程需要 5 到 11 天才能达到良好的效果。高尔基法有其特征性的缺陷,但通过这种方法,神经元和神经胶质看起来很好地被渗透,允许在光镜下进行定性和定量研究。这项技术增加了研究人类神经组织的基本技术,具有与其他物种的“单切片”高尔基法相同的优点;它简单、快速,所需设备最少,并且结果一致。

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