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评估改良高尔基-考克斯法中脑浸染时间对神经元染色及形态的影响。

Appraisal of the effect of brain impregnation duration on neuronal staining and morphology in a modified Golgi-Cox method.

作者信息

Narayanan Sareesh Naduvil, Jetti Raghu, Gorantla Vasavi Rakesh, Kumar Raju Suresh, Nayak Satheesha, Bhat P Gopalakrishna

机构信息

Department of Physiology, Melaka Manipal Medical College (Manipal Campus), Manipal University, Manipal 576 104, India.

Department of Anatomy, Melaka Manipal Medical College (Manipal Campus), Manipal University, Manipal 576 104, India.

出版信息

J Neurosci Methods. 2014 Sep 30;235:193-207. doi: 10.1016/j.jneumeth.2014.07.007. Epub 2014 Jul 23.

Abstract

BACKGROUND

Golgi-Cox staining method is considered as one of the best neurohistological and fascinating staining techniques to reveal the cytoarchitecture of the brain. Requirement of longer time (more than a month), laborious section processing steps, requirement of sophisticated equipment's and costly ready to use kits limits extensive use of this technique.

NEW METHOD

The need for a modified staining technique is to overcome some of these hurdles. Here we describe a modification of Golgi-Cox staining involving reduced impregnation time (7 days), omitting tissue dehydration steps, and alterations in section processing steps. Different impregnation duration (7 days, 14 days, 1 month, 6 months and 10 months) effects on optimized staining of dorsal hippocampus and basolateral amygdala were investigated.

RESULTS

Modified Golgi-Cox staining method was found to be effective in staining rat hippocampus and amygdala. Impregnation for 7 days, 14 days and 1 month resulted in giving good results and they were comparable. However, artifacts were slightly elevated with 6 months group but not extensively. Impregnation for 10 months negatively affected the staining process.

COMPARISON WITH EXISTING METHOD(S): Compared to existing methods the current method was found to be cost effective, fast, reliable and can be executed in labs where infrastructure is limited.

CONCLUSIONS

Current modification considerably benefitted in obtaining better results (good clarity and lesser artifact) in a short time. Longer impregnated brain sections were found to be unsuitable for morphometric evaluation due to more stain precipitation and artifact. The modified technique can be used to study cellular architecture in other brain regions.

摘要

背景

高尔基-考克斯染色法被认为是揭示大脑细胞结构的最佳神经组织学染色技术之一。该技术需要较长时间(超过一个月)、繁琐的切片处理步骤、精密设备以及昂贵的即用型试剂盒,限制了其广泛应用。

新方法

需要一种改良的染色技术来克服这些障碍。在此,我们描述了一种高尔基-考克斯染色法的改良方法,包括缩短浸染时间(7天)、省略组织脱水步骤以及改变切片处理步骤。研究了不同浸染时长(7天、14天、1个月、6个月和10个月)对背侧海马体和基底外侧杏仁核优化染色的影响。

结果

改良后的高尔基-考克斯染色法在对大鼠海马体和杏仁核进行染色时有效。7天、14天和1个月的浸染效果良好且相当。然而6个月组的伪像略有增加,但不严重。10个月的浸染对染色过程有负面影响。

与现有方法的比较

与现有方法相比,当前方法具有成本效益高、速度快、可靠且可在基础设施有限的实验室中实施的特点。

结论

当前的改良方法在短时间内获得更好的结果(清晰度高且伪像少)方面有很大益处。由于染色沉淀和伪像较多,长时间浸染的脑切片不适用于形态计量学评估。改良技术可用于研究其他脑区的细胞结构。

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