Despite a rapid growth in the application of modern techniques for visualization studies in life sciences, the classical methods of histological examination are yet to be outdated. Herein, we introduce a new approach that involves combining silver nitrate pretreatment and impregnation with consequent Nissl (cresyl violet) staining for cortex and striatum architectonics study on the same microscopy slide. The developed approach of hybrid staining provides a high-quality visualization of cellular and subcellular structures, including impregnated neurons (about 10%), Nissl-stained neurons (all the remaining ones), and astrocytes, as well as chromatophilic substances, nucleoli, and neuropil in paraffin sections. We provide a comparative study of the neuronal architectonics in both the motor cortex and striatum based on the differences in their tinctorial properties. In addition to a comparative study of the neuronal architectonics in both the motor cortex and striatum, the traditional methods to stain the cortex (motor and piriform) and the striatum are considered. The proposed staining approach compiles the routine conventional methods for thin sections, expanding avenues for more advanced examination of neurons, blood–brain barrier components, and fibers both under normal and pathological conditions. One of the main hallmarks of our method is the ability to detect changes in the number of glial cells. The results of astrocyte visualization in the motor cortex obtained by the developed technique agree well with the alternative studies by glial fibrillary acidic protein (GFAP) immunohistochemical reaction. The presented approach of combined staining has great potential in current histological practice, in particular for the evaluation of several neurological disorders in clinical, pre-clinical, or neurobiological animal studies.
|Number of pages||11|
|Journal||Frontiers in Neuroanatomy|
|Early online date||14 Oct 2022|
|Publication status||Published - 14 Oct 2022|
© 2022 Piavchenko, Soldatov, Venediktov, Kartashkina, Novikova, Gorbunova, Boronikhina, Yatskovskiy, Meglinski and Kuznetsov. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
Funding: This research was funded by the Ministry of Science and Higher Education of the Russian Federation within the framework of State support for the creation and development of World-Class Research Centers “Digital Biodesign and Personalized Healthcare” (grant number: 075-15-2022-304). IM also acknowledges partial support from the Academy of Finland
(grant projects: 351068 and 325097).
- staining combination
- Nissl staining
- silver impregnation
- protoplasmic astrocytes
- artificial pseudostaining