A combined use of silver pretreatment and impregnation with consequent Nissl staining for cortex and striatum architectonics study

Gennadii Piavchenko; Vladislav Soldatov; Artem Venediktov; Natalia Kartashkina; Natalia Novikova; Marina Gorbunova; Tatiana Boronikhina; Alexander Yatskovskiy; Igor Meglinski; Sergey Kuznetsov

doi:10.3389/fnana.2022.940993

A combined use of silver pretreatment and impregnation with consequent Nissl staining for cortex and striatum architectonics study

Front Neuroanat. 2022 Oct 14:16:940993. doi: 10.3389/fnana.2022.940993. eCollection 2022.

Affiliations

¹ Department of Histology, Cytology, and Embryology, I.M. Sechenov First Moscow State Medical University (Sechenov University), Moscow, Russia.
² Department of Pharmacology and Clinical Pharmacology, Belgorod National Research University, Belgorod, Russia.
³ Laboratory of Pathophysiology, Institute of Experimental Medicine, Saint Petersburg, Russia.
⁴ Department of Histology, Cytology, and Embryology, Orel State University named after I.S. Turgenev, Orel, Russia.
⁵ Opto-Electronics and Measurement Techniques, Faculty of Information and Electrical Engineering, University of Oulu, Oulu, Finland.
⁶ College of Engineering and Applied Science, Aston University, Birmingham, United Kingdom.

Abstract

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techniques for visualization studies in life sciences, the classical methods of histological examination are yet to be outdated.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.

Keywords: Nissl staining; artificial pseudostaining; protoplasmic astrocytes; silver impregnation; staining combination.