Advances in thin tissue Golgi-Cox impregnation: fast, reliable methods for multi-assay analyses in rodent and non-human primate brain.
Advances in thin tissue Golgi-Cox impregnation: fast, reliable methods for multi-assay analyses in rodent and non-human primate brain.
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DOI：
10.1016/j.jneumeth.2012.12.001
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发表时间：
2013-03-15
影响因子：
3
通讯作者：
Steece-Collier, Kathy
中科院分区：
文献类型：
--
作者：
Levine, Nathan D.;Rademacher, David J.;Collier, Timothy J.;O'Malley, Jennifer A.;Kells, Adrian P.;Sebastian, Waldy San;Bankiewicz, Krystof S.;Steece-Collier, Kathy
关键词：
1873年，卡米洛·高尔基发现了一种染色技术，可以对大脑中的整个神经元进行可视化，最初被称为“黑色反应”，现在被称为高尔基体受孕。尽管这种方法具有反复无常的性质，高尔基体浸渍仍然是一种广泛使用的方法，用于整个神经元的可视化以及树突树枝和脊椎的量化分析。我们描述了一系列可靠的，改进的‘高尔基-考克斯’浸渍方法，这些方法补充了一些现有的方法，并且比传统的全脑‘高尔基’浸渍方法有几个优点。首先，这些方法利用60-100μm厚的脑切片，允许快速、可靠地植入大鼠(7-14天)和非人灵长类动物(30天)的神经元，同时避免了传统上使用薄片的其他“快速高尔基体”技术的缺陷。其次，这些方法使用了几种常见的组织固定剂，导致在丙烯醛、戊二醛和多聚甲醛灌流的大鼠脑切片中以及戊二醛灌流的NHP脑组织中进行高质量的神经元浸渍。第三，因为在加工前要在振动刀上获得薄片，所以脑组织的交替切片可以用于额外的分析，如免疫组织化学或电子显微镜。这一后一优点允许例如比较相关组织化学标记或超微结构成分相邻的部分中的树枝晶形态。最后，我们描述了一种在同一组织切片中同时进行酪氨酸羟基酶免疫组织化学和高尔基体染色的光镜观察方法。因此，这里描述的方法允许快速、高质量的高尔基体受孕，并通过允许在单个动物内进行多种分析来保护实验对象。
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.
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影响因子：
3
作者：
GABBOTT, PL;SOMOGYI, J
通讯作者：
SOMOGYI, J
影响因子：
3
作者：
GLASER, EM;VANDERLOOS, H
通讯作者：
VANDERLOOS, H
影响因子：
3.2
作者：
Pilati, Nadia;Barker, Matthew;Hamann, Martine
通讯作者：
Hamann, Martine
DOI：
10.1523/jneurosci.6165-09.2010
发表时间：
2010-03-31
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Rademacher DJ;Rosenkranz JA;Morshedi MM;Sullivan EM;Meredith GE
通讯作者：
Meredith GE
影响因子：
3.7
作者：
Pinto L;Mateus-Pinheiro A;Morais M;Bessa JM;Sousa N
通讯作者：
Sousa N