A Rapid Method Combining Golgi and Nissl Staining to Study Neuronal Morphology and Cytoarchitecture

Abstract

The Golgi silver impregnation technique gives detailed information on neuronal morphology of the few neurons it labels, whereas the majority remain unstained. In contrast, the Nissl staining technique allows for consistent labeling of the whole neuronal population but gives very limited information on neuronal morphology. Most studies characterizing neuronal cell types in the context of their distribution within the tissue slice tend to use the Golgi silver impregnation technique for neuronal morphology followed by deimpregnation as a prerequisite for showing that neuron's histological location by subsequent Nissl staining. Here, we describe a rapid method combining Golgi silver impregnation with cresyl violet staining that provides a useful and simple approach to combining cellular morphology with cytoarchitecture without the need for deimpregnating the tissue. Our method allowed us to identify neurons of the facial nucleus and the supratrigeminal nucleus, as well as assessing cellular distribution within layers of the dorsal cochlear nucleus. With this method, we also have been able to directly compare morphological characteristics of neuronal somata at the dorsal cochlear nucleus when labeled with cresyl violet with those obtained with the Golgi method, and we found that cresyl violet-labeled cell bodies appear smaller at high cellular densities. Our observation suggests that cresyl violet staining is inadequate to quantify differences in soma sizes.

Cerium oxide nanoparticles promote neurogenesis and abrogate hypoxia-induced memory impairment through AMPK–PKC–CBP signaling cascade

“…Cresyl violet staining was performed as described previously 35 with slight modifications. In brief, rats were perfused with cold PBS and perfuse fixed with 4% paraformaldehyde.…”

Altered Hippocampal Gene Expression and Morphology in Fetal Piglets following Maternal Respiratory Viral Infection

“…Excess stain was rinsed in tap water, after which the slides were washed in 70% alcohol and dehydrated through absolute alcohol. The sections were cleared in xylene, mounted with DPX Mounting Medium (Sigma-Aldrich), coverslipped, and allowed to air-dry under a fume hood [49]. The exclusion criteria were (1) variable staining intensity, i.e., either too dark or too light, or (2) slide or tissue section damage.…”

Autometallographic enhancement of the Golgi-Cox staining enables high resolution visualization of dendrites and spines

“…No cell bodies were seen after 5 and 10 days of Golgi-Cox impregnation microscopy of uranyl or immunohistochemical stained sections providing superb resolution of subcellular structures although reconstruction of the dendritic arbour of a single cell is time consuming and requires rigorous sampling methodologies (Popov et al 2008). Intracellular injection of biocytin or lucifer yellow, combined with the use of confocal microscopy give likewise a nice resolution of dendrites and spines (Pilati et al 2008;Pyapali et al 1998;Ruan et al 2009;Shen et al 2008). This method will only reveal the injected cells, which may not always be Wlled completely.…”

“…New histological staining methods based on enzyme histochemistry, immunohistochemistry, and Xuorescent dyes have emerged since. However, the Golgi staining remains as a primary technique for visualization of the dendritic branching pattern and dendritic spines, because it allows isolation and visualization of the dendritic arbours from a minor random fraction of the neurons in a certain brain area (Heinz 2005;Pilati et al 2008;Rosoklija et al 2003;Spacek 1989). Accordingly, Golgi techniques are not only useful for pure anatomical studies, but also widely used in studies examining behavioural-morphological relationships (Gibb and Kolb 1998;Pilati et al 2008).…”