Definition of a sectioning plane and place for a section containing hoped-for regions using a spare counterpart specimen

Abstract

Histological examination of targets in regions of interest in histological sections is one of the most frequently used tools in biomedical research. However, it is a technical challenge to secure a multitarget section for inspection of the structure’s mutual relationship of targets or a longitudinally filamentous- or tubular-formed tissue section for visitation of the overall morphological features. We present a method with a specified cutting plane and place, allowing researchers to cut directly at the multitarget centers accurately and quickly. The method is proven to be reliable with high accuracy and reproducibility and a low coefficient of variation, testing on repeat experiments of three target’s position-known models. With this method, we successfully yielded single sections containing whole intraorbital optical nerves, three aortic valves, or whole thoracic tracheas in their central positions. The adjoined custom-made tools used in the study, such as various tissue-specific formulated calibrated trimming and embedding guides, an organ-shaped cavity plaster mold, and a two-time embedding technique for optimal and identical trimming or embedding, also bear great potential to become a common supplemental tool for traditional histology and may contribute to the reduction of the labor, and the number of animals needed.

Hypertonic saline- and detergent-accelerated EDTA-based decalcification better preserves mRNA of bones

“…Accuracy is the closeness of agreement between the tested results and accepted reference values 37 , 38 . Staining results with decalcification of 10–15% EDTA are widely used and accepted for morphological analysis.…”

Flow chamber staining modality for real-time inspection of dynamic phenotypes in multiple histological stains

“…Accuracy is the closeness of agreement between the tested results and accepted reference values [ 16 , 17 ]. Staining results with traditional (one-section-one-stain) histological procedures are widely used and accepted for morphological analysis.…”

“…The method must be reproducible and not be affected by day-to-day variation [ 16 , 17 ]. The sections from the same regions, namely the papillary muscle for the heart, the cervical segment for the esophagus, and the brain dorsal cortex at Bregma 0 mm (Figs 3B , 4B and 5B ) of native animals (6 rounds, see S1 Table in S1 File ), should return similar outcomes if reassessed at a later day with the same staining procedure.…”

Sex‐specific behavioural, metabolic, and immunohistochemical changes after repeated administration of the synthetic cannabinoid AKB48 in mice

“…Sections 8‐microns thick of both cerebellar vermis and brain hemispheres were cut in the sagittal and coronal plane, respectively, and collected on silane‐coated slides. Hence, because the CNS has more complex specialized structures compared with other tissues, a general staining method, that is, haematoxylin & eosin (H&E), was used to achieve an overview of tissue structures and anatomical order and area‐specific setting (De Luca et al, 2023; Jordan et al, 2011; Li et al, 2022; Roda et al, 2019, 2023; Xiong & Gendelman, 2014). Therefore, using the brightfield examination of H&E‐stained samples at low magnification, and based on the specific target area to be evaluated, neuroanatomical site identification was achieved, allowing the selection of precise sections of cerebellum and dorsolateral prefrontal cortex (DLPFC), which were further processed for histology and immunocytochemical analysis to gauge expected neurotoxicity (Houle, 2011).…”

“…Sections 8-microns thick of both cerebellar vermis and brain hemispheres were cut in the sagittal and coronal plane, respectively, and collected on silane-coated slides. Hence, because the CNS has more complex specialized structures compared with other tissues, a general staining method, that is, haematoxylin & eosin (H&E), was used to achieve an overview of tissue structures and anatomical order and area-specific setting (De Luca et al, 2023;Jordan et al, 2011;Li et al, 2022;Roda et al, 2019Roda et al, , 2023Xiong & Gendelman, 2014).…”