Michigan State University, College of Human Medicine, Department of Translational Science and Molecular Medicine, 333 Bostwick Avenue NE, Grand Rapids, MI 49503, USA.
J Neurosci Methods. 2013 Mar 15;213(2):214-27. doi: 10.1016/j.jneumeth.2012.12.001. Epub 2013 Jan 8.
In 1873 Camillo Golgi discovered a staining technique that allowed for the visualization of whole neurons within the brain, initially termed 'the black reaction' and is now known as Golgi impregnation. Despite the capricious nature of this method, Golgi impregnation remains a widely used method for whole neuron visualization and analysis of dendritic arborization and spine quantification. We describe a series of reliable, modified 'Golgi-Cox' impregnation methods that complement some existing methods and have several advantages over traditional whole brain 'Golgi' impregnation. First, these methods utilize 60-100μm thick brain sections, which allows for fast, reliable impregnation of neurons in rats (7-14 days) and non-human primates (NHP) (30 days) while avoiding the pitfalls of other 'rapid Golgi' techniques traditionally employed with thin sections. Second, these methods employ several common tissue fixatives, resulting in high quality neuron impregnation in brain sections from acrolein, glutaraldehyde, and paraformaldehyde perfused rats, and in glutaraldehyde perfused NHP brain tissue. Third, because thin sections are obtained on a vibratome prior to processing, alternate sections of brain tissue can be used for additional analyses such as immunohistochemistry or electron microscopy. This later advantage allows for comparison of, for example, dendrite morphology in sections adjacent to pertinent histochemical markers or ultrastructural components. Finally, we describe a method for simultaneous light microscopic visualization of both tyrosine hydroxylase immunohistochemistry and Golgi impregnation in the same tissue section. Thus, the methods described here allow for fast, high quality Golgi impregnation and conserve experimental subjects by allowing multiple analyses within an individual animal.
1873 年，Camillo Golgi 发现了一种染色技术，可以在大脑中可视化整个神经元，最初称为“黑色反应”，现在称为 Golgi 浸渍。尽管这种方法具有多变性，但 Golgi 浸渍仍然是一种广泛使用的方法，用于可视化整个神经元和分析树突分支和棘突的量化。我们描述了一系列可靠的、经过改良的“Golgi-Cox”浸渍方法，这些方法补充了一些现有的方法，并且相对于传统的全脑“Golgi”浸渍具有几个优势。首先，这些方法使用 60-100μm 厚的脑切片，这允许在大鼠（7-14 天）和非人类灵长类动物（NHP）（30 天）中快速、可靠地浸渍神经元，同时避免了传统上使用薄切片的其他“快速 Golgi”技术的陷阱。其次，这些方法采用了几种常见的组织固定剂，从而在大鼠经丙烯醛、戊二醛和多聚甲醛灌注以及 NHP 脑组织经戊二醛灌注的脑切片中实现了高质量的神经元浸渍。第三，由于在处理前在振动切片机上获得了薄切片，因此可以对脑组织的交替切片进行其他分析，例如免疫组织化学或电子显微镜。这一后来的优势允许例如在与相关组织化学标记物或超微结构成分相邻的切片中比较树突形态。最后，我们描述了一种在同一组织切片中同时可视化酪氨酸羟化酶免疫组织化学和 Golgi 浸渍的方法。因此，这里描述的方法允许快速、高质量的 Golgi 浸渍，并通过允许在单个动物内进行多种分析来节省实验对象。
