B Landzhov, D Hinova-Palova, K Fakih, L Edelstein, L Gaydarski, A Alexandrov, V Kirkov, A Paloff, E Radeva
{"title":"Corticoclaustral connections in the cat.","authors":"B Landzhov, D Hinova-Palova, K Fakih, L Edelstein, L Gaydarski, A Alexandrov, V Kirkov, A Paloff, E Radeva","doi":"10.1080/01478885.2025.2476835","DOIUrl":"https://doi.org/10.1080/01478885.2025.2476835","url":null,"abstract":"<p><p>The claustrum is a sheet-like layer of gray matter situated between the external and extreme capsules of the mammalian brain. This structure was first described by the French physician and anatomist Vicq d'Azyr in 1786. The claustrum's phylogeny, ontogeny and functional characteristics have long been the subject of debate and considerable investigative efforts. However, despite such efforts (or perhaps as a result thereof), significant disparities and discrepancies remain, most notably in the context of the claustrum's afferent and efferent connections. For the purpose of this study, we sought to focus our efforts on fronto-claustral and occipito-claustral connections. Twelve healthy, adult male cats, each weighing ~ 3.5 kg, were studied, seven of which underwent electrolytic lesions of the frontal cortex (A3, A4, and a portion of A6), and five of the occipital cortex (A17, A18, A21). From three to six days after lesioning, subjects were euthanized in accordance with ethical norms. After the brains were removed and blocked, samples of the claustrum were taken and prepared for electron microscopy. Three to six days after lesions of the frontal cortex, we observed an abundance of degenerative boutons in the dorsal claustrum. The vast majority of boutons exhibited the characteristic signs of dark degeneration, whereas only 10% appeared to have undergone light degeneration. Similar results were seen in the dorsal claustrum over the same period of time following lesions of the visual cortex. These results suggest that the dorsal claustrum receives at least two types of connections - separately and concurrently - from the frontal and occipital cortices.</p>","PeriodicalId":15966,"journal":{"name":"Journal of Histotechnology","volume":" ","pages":"1-11"},"PeriodicalIF":0.6,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143624935","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Charles Havnar, Loryn Holokai, Ryan Ichikawa, Wennie Chen, Alexis Scherl, Eliah R Shamir
{"title":"Histogel-based techniques for embedding organoids in paraffin blocks enable high throughput downstream histopathological analyses.","authors":"Charles Havnar, Loryn Holokai, Ryan Ichikawa, Wennie Chen, Alexis Scherl, Eliah R Shamir","doi":"10.1080/01478885.2024.2398381","DOIUrl":"10.1080/01478885.2024.2398381","url":null,"abstract":"<p><p>Organoids are <i>in vitro</i> tissue models derived from human or animal primary tissues or stem cells that allow for studying three-dimensional (3D) tissue biology, toxicity testing, biomarker evaluation, and assessment of compound efficacy, supplementing or potentially minimizing use of animal models. Organoids are typically cultured in a 3D format within an extracellular matrix and, at the end of an experiment, can be further processed for various cellular or molecular readouts. Analysis often relies on whole mount immunolabeling for markers of interest, which consumes the entire sample/well, thereby limiting sample availability for downstream assays. In addition, 3D cultures become more friable after fixation and are susceptible to sample loss during washing steps. In contrast, by fixing and processing organoids to a paraffin block, dozens or hundreds of unstained slides can be generated, enabling robust characterization via multiple assays, including histologic evaluation and (immuno)histochemical stains, thus maximizing the yield of these time- and labor-intensive cultures. Here we describe three methods to process 3D Matrigel cultures into paraffin blocks using Histogel as an embedding agent. The three techniques all yield high-quality sections but vary in complexity of implementation at different steps, and their application for different use cases is discussed.</p>","PeriodicalId":15966,"journal":{"name":"Journal of Histotechnology","volume":" ","pages":"46-57"},"PeriodicalIF":0.6,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142132950","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sheila Criswell, Brian Gaylord, Christopher R Pitzer
{"title":"Histological methods for plant tissues.","authors":"Sheila Criswell, Brian Gaylord, Christopher R Pitzer","doi":"10.1080/01478885.2024.2397989","DOIUrl":"10.1080/01478885.2024.2397989","url":null,"abstract":"<p><p>Although many of the structures and organelles of vegetative cells are comparable to those of animal tissues, significant differences between the two kingdoms require modifications in histological techniques for both tissue processing steps and histochemical staining techniques. The authors investigated the challenges of working with plant tissues by collecting various flora to represent the four main plant organs: leaf, stem, root, and flower/fruit. Triplicate samples for each specimen were placed into formalin for paraffin embedding, placed into formalin for later frozen sections, and used fresh to undergo immediate frozen sectioning. Frozen sections of plant tissues were more difficult to obtain than formalin-fixed paraffin-embedded (FFPE) sections, exhibited tissue loss during staining, and were inferior morphologically to FFPE sections. Although, historically, plant tissue fixation and processing has employed several different reagents compared with those used in animal tissue processing and took significantly longer times, the current investigation determined reagents and protocols from a modern histology laboratory which processes mammalian tissues can be applied to plant tissue processing with only slight modifications in respect to reagent timing. Additionally, staining techniques were compared and while it is well known that plant cell walls stain well with safranin O, the current investigation determined the uptake of safranin O can be accelerated by incubating at 60°C.</p>","PeriodicalId":15966,"journal":{"name":"Journal of Histotechnology","volume":" ","pages":"58-67"},"PeriodicalIF":0.6,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142120024","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Resolving the bone - optimizing decalcification in spatial transcriptomics and molecular pathology.","authors":"Shuoshuo Wang","doi":"10.1080/01478885.2024.2446038","DOIUrl":"10.1080/01478885.2024.2446038","url":null,"abstract":"<p><p>Bone tissue poses critical roadblocks for spatial transcriptomics and molecular pathology due to a combination of its dense, calcified matrix and inadequate preservation of biomolecules in conventional decalcification. Decalcification is a complex and nuanced histological process to concomitantly preserve nucleic acids, proteins, and tissue architecture, ensuring molecular integrity for downstream assays. However, commonly used agents like formic and hydrochloric acids, while efficient, can degrade biomolecules to varying extents, complicating assays such as PCR, sequencing, immunohistochemistry, and <i>in situ</i> hybridization. Advances in spatial transcriptomics, both sequencing- and imaging-based, emphasize the importance of optimizing decalcification protocols to improve research outcomes. This synoptic and perspective article explores traditional and modern decalcification methods, offering recommendations on technical and methodological refinements for achieving molecularly robust processing of bone and calcified tissues in spatial transcriptomics and molecular pathology.</p>","PeriodicalId":15966,"journal":{"name":"Journal of Histotechnology","volume":" ","pages":"68-77"},"PeriodicalIF":0.6,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142895270","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
David Krull, Premi Haynes, Anil Kesarwani, Julien Tessier, Benjamin J Chen, Kelly Hunter, Deniliz Rodriguez, Yan Liang, Jim Mansfield, Maxine McClain, Corinne Ramos, Edward Bonnevie, Esperanza Anguiano
{"title":"A best practices framework for spatial biology studies in drug discovery and development: enabling successful cohort studies using digital spatial profiling.","authors":"David Krull, Premi Haynes, Anil Kesarwani, Julien Tessier, Benjamin J Chen, Kelly Hunter, Deniliz Rodriguez, Yan Liang, Jim Mansfield, Maxine McClain, Corinne Ramos, Edward Bonnevie, Esperanza Anguiano","doi":"10.1080/01478885.2024.2391683","DOIUrl":"10.1080/01478885.2024.2391683","url":null,"abstract":"<p><p>The discovery of biomarkers, essential for successful drug development, is often hindered by the limited availability of tissue samples, typically obtained through core needle biopsies. Standard 'omics platforms can consume significant amounts of tissue, forcing scientist to trade off spatial context for high-plex assays, such as genome-wide assays. While bulk gene expression approaches and standard single-cell transcriptomics have been valuable in defining various molecular and cellular mechanisms, they do not retain spatial context. As such, they have limited power in resolving tissue heterogeneity and cell-cell interactions. Current spatial transcriptomics platforms offer limited transcriptome coverage and have low throughput, restricting the number of samples that can be analyzed daily or even weekly. While the Digital Spatial Profiling (DSP) method does not provide single-cell resolution, it presents a significant advancement by enabling scalable whole transcriptome and ultrahigh-plex protein analysis from distinct tissue compartments and structures using a single tissue slide. These capabilities overcome significant constraints in biomarker analysis in solid tissue specimens. These advancements in tissue profiling play a crucial role in deepening our understanding of disease biology and in identifying potential therapeutic targets and biomarkers. To enhance the use of spatial biology tools in drug discovery and development, the DSP Scientific Consortium has created best practices guidelines. These guidelines, built on digital spatial profiling data and expertise, offer a practical framework for designing spatial studies and using current and future spatial biology platforms. The aim is to improve tissue analysis in all research areas supporting drug discovery and development.</p>","PeriodicalId":15966,"journal":{"name":"Journal of Histotechnology","volume":" ","pages":"7-26"},"PeriodicalIF":0.6,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142120015","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Carolyn Dunlap, Niky Zhao, Linda S Ertl, Thomas J Schall, Kathleen M C Sullivan
{"title":"C5aR expression in kidney tubules, macrophages and fibrosis.","authors":"Carolyn Dunlap, Niky Zhao, Linda S Ertl, Thomas J Schall, Kathleen M C Sullivan","doi":"10.1080/01478885.2024.2430041","DOIUrl":"10.1080/01478885.2024.2430041","url":null,"abstract":"<p><p>The anaphylatoxin C5a and its receptor C5aR (CD88) are complement pathway effectors implicated in renal diseases, including ANCA-associated vasculitis. We investigated the kidney expression of C5aR and a second C5a receptor C5L2 by using immunohistochemistry, in situ hybridization, and spatial gene expression on formalin-fixed, paraffin-embedded human and mouse kidney. C5aR was detected on interstitial macrophages and in multiple tubular regions, including distal and proximal; C5L2 had a similar expression pattern. The 5/6 nephrectomy model of chronic kidney injury exhibited increased C5aR expression by infiltrating cells within the fibrotic regions. C5aR expression was confirmed on human leukocytes and in vitro differentiated macrophages by flow cytometry, and treatment with C5a induced the expression of chemokines and remodeling factors by macrophages, including CCL-3/-4/-7, -20, MMP-1/-3/-8/-12, and F3, and promoted leukocyte survival. C5a activity was C5aR dependent, as demonstrated by reversal with the C5aR inhibitor avacopan. Collectively, these results suggest that myeloid C5aR may induce excessive inflammation in the kidney via immune cell recruitment, extracellular matrix destruction, and remodeling, resulting in fibrotic tissue deposition.</p>","PeriodicalId":15966,"journal":{"name":"Journal of Histotechnology","volume":" ","pages":"27-45"},"PeriodicalIF":0.6,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142739616","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Histotechnology at the crossroad of spatial biology: bridging legacy and innovation.","authors":"Shuoshuo Wang, Yongfu Wang, David Krull","doi":"10.1080/01478885.2025.2456415","DOIUrl":"10.1080/01478885.2025.2456415","url":null,"abstract":"","PeriodicalId":15966,"journal":{"name":"Journal of Histotechnology","volume":" ","pages":"1-6"},"PeriodicalIF":0.6,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143189498","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Jeffery T Williams, Tracy A Goodpaster, Miki Haraguchi
{"title":"Optimizing tissue adherence on glass slides using polyurethane glue: a new slide preparation method.","authors":"Jeffery T Williams, Tracy A Goodpaster, Miki Haraguchi","doi":"10.1080/01478885.2024.2440679","DOIUrl":"https://doi.org/10.1080/01478885.2024.2440679","url":null,"abstract":"<p><p>The application of Clear Gorilla Glue® household adhesive to microscope slides has been found to enhance the mounting and retention of traditionally difficult tissue types, notably bone and tooth specimens. Improvement in end results were observed across H&E staining, immunohistochemistry, and <i>in situ</i> hybridization.</p>","PeriodicalId":15966,"journal":{"name":"Journal of Histotechnology","volume":" ","pages":"1-3"},"PeriodicalIF":0.6,"publicationDate":"2024-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142837135","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Multiple levels of Gomori methenamine silver (GMS) stains do not improve diagnostic yield in esophageal biopsies.","authors":"Michael Occidental, Andrew Dunn, Aaron R Huber","doi":"10.1080/01478885.2024.2437587","DOIUrl":"https://doi.org/10.1080/01478885.2024.2437587","url":null,"abstract":"<p><p>Gomori methenamine silver (GMS) stains are commonly utilized to exclude fungal esophagitis in esophageal biopsies. Our laboratory protocol for special stains, including GMS, stipulates that two levels be performed for each stain. We retrospectively reviewed 127 esophageal biopsies which had GMS stains performed; there were 62 GMS-positive and 65 GMS-negative cases. Sixty-one of the positive cases (98%) had fungal organisms present on all levels. Only one case showed fungal elements on the first, but not the second, level. Given national laboratory staff shortages, performing levels for special stains may be unnecessary, inefficient, and time consuming in the surgical pathology laboratory.</p>","PeriodicalId":15966,"journal":{"name":"Journal of Histotechnology","volume":" ","pages":"1-3"},"PeriodicalIF":0.6,"publicationDate":"2024-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142813158","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nathan T Sheppard, Melissa C Daniel, Noah S Nelson, Alexis Donneys, Steven R Buchman
{"title":"Optimizing immunofluorescent staining of H vessels within an irradiated fracture callus in paraffin-embedded tissue samples.","authors":"Nathan T Sheppard, Melissa C Daniel, Noah S Nelson, Alexis Donneys, Steven R Buchman","doi":"10.1080/01478885.2024.2371060","DOIUrl":"10.1080/01478885.2024.2371060","url":null,"abstract":"<p><p>H vessels are an essential link in angiogenic-osteogenic coupling and orchestrate the process of bone healing. H vessels are critically deficient in the setting of radiation-induced fractures, which have been reported to occur in up to 25% of patients undergoing radiotherapy. By increasing H-vessel proliferation, Deferoxamine (DFO) revitalizes the physiologic response to skeletal injury and accelerates irradiated fracture repair. H-vessel quantification is therefore an important outcome measure in histologic analysis of bone healing. However, an optimized protocol for staining H vessels in formalin-fixed paraffin-embedded (FFPE) tissue sections has not been reported. With this protocol, we describe a method of staining FFPE bone samples with minimal background fluorescence and high signal-to-noise ratio. We examined mandibular specimens in a rat model of bone healing from a range of fracture conditions, including healthy bone (Fx), irradiated bone (XFx), and irradiated bone with DFO treatment (XFx-DFO). Quantitative analysis revealed a significant increase of H vessels in the XFxDFO group compared to both the Fx and XFx groups. By optimizing immunofluorescent staining of H vessels in FFPE samples across a range of fracture conditions, we offer investigators an efficacious means of producing reliable imaging for quantitative analysis of H vessels in an irradiated fracture callus.</p>","PeriodicalId":15966,"journal":{"name":"Journal of Histotechnology","volume":" ","pages":"173-179"},"PeriodicalIF":0.6,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141492250","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}