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Optimising Golgi-Cox staining for use with perfusion-fixed brain tissue validated in the zQ175 mouse model of Huntington's disease

Bayram-Weston, Zubeyde, Olsen, Elliott, Harrison, David, Dunnett, Stephen Bruce and Brooks, Simon 2016. Optimising Golgi-Cox staining for use with perfusion-fixed brain tissue validated in the zQ175 mouse model of Huntington's disease. Journal of Neuroscience Methods 265 , pp. 81-88. 10.1016/j.jneumeth.2015.09.033

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Abstract

Background: The Golgi-Cox stain is an established method for characterising neuron cell morphology. The method highlights neurite processes of stained cells allowing the complexity of dendritic branching to be measured. New Method: Conventional rapid Golgi and Golgi-Cox methods all require fresh impregnation in unfixed brain blocks. Here, we describe a modified method that gives high quality staining on brain tissue blocks perfusion-fixed with 4% paraformaldehyde (PFA) and post-fixed by immersion for 24 hr. Results: Tissue perfused with 4% PFA and post fixed for 24 hours remained viable for the modified Golgi-Cox silver impregnation staining of mouse striatum from perfused wild type and zQ175. It was not found necessary to impregnate tissue blocks with Golgi solutions prior to sectioning, as post-sectioned tissues yielded equally good impregnation. Impregnation for 14 days resulted in optimal visualization of striatal neuron and dendritic morphology. Although no modifications applied to the Rapid Golgi method were reliable, the modified Golgi-Cox method yielded consistently reliable high-quality staining. Comparison with Existing Methods: The current method used fixed tissues to reduce damage and preserve cell morphology. The revised method was found to be fast, reliable and cost effective without the need for expensive staining kits and could be performed in any neuroscience lab with limited specialist equipment. Conclusions: The present study introduces a robust reproducible and inexpensive staining method for identifying neuronal morphological changes in the post fixed mouse brain, and is suitable for assessing changes in cell morphology in models of neurodegeneration and in response to experimental treatment.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Biosciences
Subjects: R Medicine > R Medicine (General)
Publisher: Elsevier
ISSN: 0165-0270
Funders: MRC
Date of First Compliant Deposit: 30 March 2016
Date of Acceptance: 30 September 2015
Last Modified: 24 Dec 2017 20:21
URI: http://orca.cf.ac.uk/id/eprint/78619

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