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

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techniques for visualization studies in life sciences, the classical methods of histological examination are yet to be outdated.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

The number of the intraepithelial T cells correlate with the proliferation index in human bulbourethral gland epithelium

Background: Our study has immunohistochemically examined T cells localization and number as well as proliferative activity level for the bulbourethral gland epithelium in men of different ages, using monoclonal antibodies against CD45RO and proliferating cell nuclear antigen (PCNA). Results: We have found that the T cells have been localized mainly in excretory ducts epithelium of the glands in any age group, meanwhile their relative number varies with age. The excretory ducts epithelium has shown a high proliferative activity when in acini the PCNA index has been low. Postnatal dynamics of the epithelium proliferative activity positively correlates with age-related density fluctuations in lymphocytic infiltration of the glands. Conclusions: We consider that intraepithelial T cells may contribute to the regulation of epithelial cells proliferation in the bulbourethral glands

Embossed topographic depolarisation maps of biological tissues with different morphological structures

Layered topographic maps of the depolarisation due to diffuse biological tissues are produced using a polarisation-holographic Mueller matrix method approach. Histological sections of myocardial tissue with a spatially structured optically anisotropic fibrillar network, and parenchymal liver tissue with a polycrystalline island structure are successfully mapped. The topography of the myocardium maps relates to the scattering multiplicity within the volume and the specific morphological structures of the biological crystallite networks. The overall depolarisation map is a convolution of the effects of these two factors. Parenchymal liver tissues behave broadly similarly, but the different biological structures present cause the degree of scattering multiplicity to increase more rapidly with increasing phase. Through statistical analysis, the dependences of the magnitudes of the first to fourth order statistical moments are determined. These moments characterise the changing distributions of the depolarisation values through the volume of biological tissues with different morphological structures. Parenchymal liver tissue depolarisation maps are characterised by larger mean and variance, and less skewness and kurtosis, compared to the distributions for the myocardium. This work demonstrates that a polarisation-holographic Mueller matrix method can be applied to the assessment of the 3D morphology of biological tissues, with applications in disease diagnosis

The number of the intraepithelial T cells correlate with the proliferation index in human bulbourethral gland epithelium

Abstract Background: Our study has immunohistochemically examined T cells localization and number as well as proliferative activity level for the bulbourethral gland epithelium in men of different ages, using monoclonal antibodies against CD45RO and proliferating cell nuclear antigen (PCNA). Results: We have found that the T cells have been localized mainly in excretory ducts epithelium of the glands in any age group, meanwhile their relative number varies with age. The excretory ducts epithelium has shown a high proliferative activity when in acini the PCNA index has been low. Postnatal dynamics of the epithelium proliferative activity positively correlates with age-related density fluctuations in lymphocytic infiltration of the glands. Conclusions: We consider that intraepithelial T cells may contribute to the regulation of epithelial cells proliferation in the bulbourethral glands

Structural changes of erythrocyte membrane revealed by 3D confocal optical profilometer

We examined hematological changes influenced by the experimental hypervitaminosis A. The 3D confocal optical profilometer was applied for assessment of the erythrocytes’ membrane structural changes influenced by an overdose of vitamin A. The blood smears were evaluated in terms of alterations of geometrical and optical parameters of erythrocytes for two groups of animals: oil base and retinol palmitate (n=9 animals for each group). The results demonstrate that an overdose of retinol palmitate causes changes in the torus curvature and pallor of discocytes, their surface area and volume. The observed structural malformations of the shape of red blood cells become visible at the earlier pre-clinical stage of changes in animal state and behaviour. With this in mind, the results of the study open a new area of research in the certain dysfunction diagnosis of red blood cells and has a great potential in the further development of new curative protocols

Digital histo-biophotonics:embossed topographic depolarization mapping of tissue samples

Abstract New Mueller matrix-based embossed depolarization mapping is developed to assess quantitatively morphological biotissues abnormalities in 3D. Typical results demonstrating polarization property variations for cardiac and liver tissues are presented

Structural changes of erythrocyte membrane revealed by 3D confocal optical profilometer

Abstract We examined hematological changes influenced by the experimental hypervitaminosis A. The 3D confocal optical profilometer was applied for assessment of the erythrocytes’ membrane structural changes influenced by an overdose of vitamin A. The blood smears were evaluated in terms of alterations of geometrical and optical parameters of erythrocytes for two groups of animals: oil base and retinol palmitate (n = 9 animals for each group). The results demonstrate that an overdose of retinol palmitate causes changes in the torus curvature and pallor of discocytes, their surface area and volume. The observed structural malformations of the shape of red blood cells become visible at the earlier preclinical stage of changes in animal state and behavior. With this in mind, the results of the study open a new area of research in the certain dysfunction diagnosis of red blood cells and have a great potential in the further development of new curative protocols