A refined Golgi-Cox method for the staining of embryonic neurons in the mouse brain

IF 2.7 4区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Neuroscience Methods Pub Date : 2025-03-19 DOI:10.1016/j.jneumeth.2025.110432

Benjamin Mitchell , Cooper Atterton , Darryl Whitehead , Stefan Thor , Michael Piper

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Abstract

The Golgi-Cox stain remains a valuable technique used to investigate the morphology of individual neurons. Despite this, Golgi-Cox staining protocols are predominantly designed to impregnate adult neurons. Protocols optimised for the staining of immature embryonic mouse neurons have been previously developed but have limitations, including being time-consuming and being reliant on the use of expensive commercial kits. Here, we present a simple and inexpensive method for Golgi-Cox staining of embryonic neurons in the mouse brain. We identified that a 60 minute, 4 % paraformaldehyde (PFA) brain fixation step, followed by a wash with distilled water prior to immersion in Golgi-Cox solution was critical to the success of the stain. By altering the duration of the wash step, the visualisation of different populations across the neuraxis of neurons could be emphasised. Shorter washes enabled cortical neurons to be readily distinguished, whereas extending the wash steps was needed to enable subcortical neurons to be delineated.

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一种用于小鼠大脑胚胎神经元染色的改进高尔基-考克斯染色法。

高尔基-考克斯染色仍然是一种有价值的技术，用于研究单个神经元的形态。尽管如此，高尔基-考克斯染色方案主要设计用于浸渍成年神经元。之前已经开发了用于未成熟胚胎小鼠神经元染色的优化方案，但存在局限性，包括耗时和依赖于使用昂贵的商业试剂盒。在这里，我们提出了一种简单而廉价的方法，用于小鼠大脑胚胎神经元的高尔基-考克斯染色。我们发现，60分钟的4%多聚甲醛（PFA）脑固定步骤，然后在浸泡在高尔基-考克斯溶液中之前用蒸馏水清洗是染色成功的关键。通过改变洗涤步骤的持续时间，可以强调神经元神经轴上不同种群的可视化。较短的洗涤时间可以很容易地区分皮层神经元，而延长洗涤时间则可以区分皮层下神经元。

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来源期刊

Journal of Neuroscience Methods 医学-神经科学

CiteScore

7.10

自引率

3.30%

发文量

226

审稿时长

52 days

期刊介绍： The Journal of Neuroscience Methods publishes papers that describe new methods that are specifically for neuroscience research conducted in invertebrates, vertebrates or in man. Major methodological improvements or important refinements of established neuroscience methods are also considered for publication. The Journal''s Scope includes all aspects of contemporary neuroscience research, including anatomical, behavioural, biochemical, cellular, computational, molecular, invasive and non-invasive imaging, optogenetic, and physiological research investigations.