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Determining factors for optimal neuronal and glial Golgi-Cox staining
Ist Teil von
Histochemistry and cell biology, 2020-10, Vol.154 (4), p.431-448
Ort / Verlag
Berlin/Heidelberg: Springer Berlin Heidelberg
Erscheinungsjahr
2020
Quelle
MEDLINE
Beschreibungen/Notizen
Golgi staining allows for the analysis of neuronal arborisations and connections and is considered a powerful tool in basic and clinical neuroscience. The fundamental rules for improving neuronal staining using the
Golgi-Cox
method are not fully understood; both intrinsic and extrinsic factors may control the staining process. Therefore, various conditions were tested to improve the
Golgi-Cox
protocol for vibratome-cut rat brain sections. Optimal staining of cortical neurons was achieved after 72 h of impregnation. Well-stained neurons in both cortical and subcortical structures were observed after 96 h of impregnation. The dendritic arborisation pattern of cortical neurons derived from the 72-h impregnation group was comparable to those of the 96 and 168-h impregnation groups. The entire brain was stained well when the pH of the
Golgi-Cox
solution was 6.5 and that of the sodium carbonate solution was 11.2. Lack of brain perfusion or perfusion with 0.9% NaCl did not influence optimal neuronal staining. Perfusion with 37% formaldehyde, followed by impregnation, only resulted in glial staining, but perfusion with 4% formaldehyde facilitated both glial and neuronal staining. Whole brains required longer impregnation times for better staining. Although every factor had a role in determining optimal neuronal staining, impregnation time and the pH of staining solutions were key factors among them. This modified
Golgi-Cox
protocol provides a simple and economical procedure to stain both neurons and glia separately.