{"title":"The impact of gliomas on the normal brain microenvironment: a pilot study.","authors":"Nicole Riha, Jacen S Moore, Sheila Criswell","doi":"10.1080/01478885.2024.2408505","DOIUrl":"https://doi.org/10.1080/01478885.2024.2408505","url":null,"abstract":"<p><p>Gliomas are malignant tumors of neuronal support cells within the central nervous system (CNS) and are characterized by poor overall prognoses and limited treatment options due to their infiltrative growth patterns. The neural tumor microenvironment, composed of benign neurons, neuroglia, endothelial cells, and intravascular white blood cells, is a target-rich site for potential chemotherapeutic agents. This study assessed cell proliferation rates, white blood cell components, and a limited number of nuclear, cytoplasmic, and membrane markers using immunohistochemistry (IHC) assays on formalin-fixed and paraffin-embedded benign and glial tumor tissue samples from the CNS. It was observed that glioma tissues had increased rates of glial cell proliferation and significant increases in the number of observed T-lymphocytes and granulocytes but decreased expression of markers Somatostatin receptor 2 (SSTR2), L1 cell adhesion molecule (L1CAM), and GATA binding protein 3 (GATA3) when compared to benign tissue samples. Understanding the lack of protein expression and population expansion potential of the glioma microenvironment in greater detail could help identify valuable therapeutic target combinations for future treatments.</p>","PeriodicalId":15966,"journal":{"name":"Journal of Histotechnology","volume":" ","pages":"1-10"},"PeriodicalIF":0.6,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142348107","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":"Adaptation of the HistoEnder, an open-source 3D printer for automated transmission electron microscopy grid staining.","authors":"Philip Seifert","doi":"10.1080/01478885.2024.2403872","DOIUrl":"10.1080/01478885.2024.2403872","url":null,"abstract":"<p><p>The HistoEnder, an inexpensive open-source 3D printer published as an automated histological slide stainer, has been adapted for conventional biological transmission electron microscopy (TEM) batch grid staining. Details are presented of the 3D printed apparatus, assembly, G-code programming, and operation on the 3D printer to post-section stains up to 20 grids through aqueous uranyl acetate, distilled water rinses, and lead stains. TEM Results are identical to manual staining with the advantages of automation using the low cost HistoEnder, apparatus, and equipment.</p>","PeriodicalId":15966,"journal":{"name":"Journal of Histotechnology","volume":" ","pages":"1-5"},"PeriodicalIF":0.6,"publicationDate":"2024-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142289123","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}
Joseph A. Esposito, Camryn J. Vader, Joel R. Israel, Steve A. McClain
{"title":"A study of PGP 9.5 immunohistochemical labeling on formalin-fixed paraffin embedded tissues for epidermal nerve fiber density testing","authors":"Joseph A. Esposito, Camryn J. Vader, Joel R. Israel, Steve A. McClain","doi":"10.1080/01478885.2024.2405418","DOIUrl":"https://doi.org/10.1080/01478885.2024.2405418","url":null,"abstract":"There have been several methods established for immunohistochemical labeling of the PGP 9.5 antigen in human tissue for the assessment of epidermal nerve fiber density, none of which uses neutral-b...","PeriodicalId":15966,"journal":{"name":"Journal of Histotechnology","volume":"19 1","pages":""},"PeriodicalIF":1.1,"publicationDate":"2024-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142261097","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":"https://doi.org/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":"1-12"},"PeriodicalIF":0.6,"publicationDate":"2024-09-05","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}
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":"https://doi.org/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":"1-20"},"PeriodicalIF":0.6,"publicationDate":"2024-09-03","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}
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":"https://doi.org/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":"1-10"},"PeriodicalIF":0.6,"publicationDate":"2024-09-02","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":"Survey of NAACLS accredited histotechnology programs in the United States.","authors":"Carol Bain, Debra Wood, Sheila Criswell","doi":"10.1080/01478885.2024.2327095","DOIUrl":"10.1080/01478885.2024.2327095","url":null,"abstract":"<p><p>Histotechnology educational programs are accredited by the National Accrediting Agency for Clinical Laboratory Sciences (NAACLS) and currently number fewer than 50 in the United States which contributes to the shortages of laboratory personnel. A survey tool designed with REDCap software was distributed to all programs identified on the NAACLS website and consisted of three parts: a) program information, b) facility information, and c) challenges. Programs are located primarily in large urban centers where populations are most concentrated. The median class size was 6 which may explain the excellent student outcomes to include 96% graduation rates and 90.7% board of registry examination pass rates. Overall, programs had ample equipment, funding, and administrative support. Costs to attend the programs were relatively low (<$3,000 per semester) for over half of the programs. However, due to the small number of accredited education programs across the US, potential students do not often have access to an institution in their area. The programs indicated that the most common challenge was recruitment of adequate high-quality candidates which may explain, in part, the persistent shortage of personnel in the histology laboratory.</p>","PeriodicalId":15966,"journal":{"name":"Journal of Histotechnology","volume":" ","pages":"109-116"},"PeriodicalIF":0.6,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140143574","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}
Dibyajyoti Boruah, A W Kashif, Barun Kumar Chakrabarty, Sarika Harikrishnan, Arijit Sen
{"title":"Correlation of light and electron microscopic morphometric parameters of glomerular capillaries with serum creatinine and proteinuria.","authors":"Dibyajyoti Boruah, A W Kashif, Barun Kumar Chakrabarty, Sarika Harikrishnan, Arijit Sen","doi":"10.1080/01478885.2024.2326274","DOIUrl":"10.1080/01478885.2024.2326274","url":null,"abstract":"<p><p>Waste products in the bloodstream are filtered by the glomerular capillaries in the kidneys and excreted into the urine. When making a differential diagnosis of kidney diseases, structural assessment of glomeruli using histological, ultrastructural, and immunological studies is crucial. This study assessed the microscopic and ultrastructural morphometric parameters of glomerular capillaries and examined their correlation with serum creatinine and proteinuria. A total of 60 kidney biopsy cases received by the transmission electron microscope (TEM) laboratory for diagnosis were included in the study. Toluidine blue stained 300 nm thick sections of TEM tissue blocks were scanned for glomerular morphometry by a whole slide imaging system, and the estimation of Bowman's capsule (BC) area, glomerular capillary lumen diameter (GCLD), glomerular capillary density (GCD), glomerular capillary surface area density (GCSA), and percentage of glomerular capillary lumen space (%GCLS) was performed with QuPath software. TEM images of 70 nm thick sections were used for the evaluation of endothelial fenestration diameter (EFD), glomerular basement membrane (GBM) thickness, and podocyte foot process (PFP) effacement. Proteinuria and serum creatinine showed positive correlations with GBM thickness and PFP effacement. Negative correlations of serum creatinine were observed with EFD, %GCLS, and GCSA. Hence, glomerular filtration is greatly affected by the total area of the glomerular capillary surface and structural changes of GBM. Reduction of glomerulus filtration due to foot process effacement and thickening of GBM results in damage to the filtration barrier leading to the leakage of plasma protein into urine.</p>","PeriodicalId":15966,"journal":{"name":"Journal of Histotechnology","volume":" ","pages":"97-108"},"PeriodicalIF":0.6,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140094188","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}
Dimka Hinova-Palova, Boycho Landzhov, Lawrence Edelstein, Khodor Fakih, Alexandar Alexandrov, Teodora Kiriakova, Elka Radeva, Lyubomir Gaydarski, Frank Denaro, Adrian Paloff
{"title":"Identification of degenerated synaptic boutons in the dorsal claustrum of the cat after electrolytic lesions of the intralaminar thalamic nuclei.","authors":"Dimka Hinova-Palova, Boycho Landzhov, Lawrence Edelstein, Khodor Fakih, Alexandar Alexandrov, Teodora Kiriakova, Elka Radeva, Lyubomir Gaydarski, Frank Denaro, Adrian Paloff","doi":"10.1080/01478885.2024.2335827","DOIUrl":"10.1080/01478885.2024.2335827","url":null,"abstract":"<p><p>The aim of this study is to investigate whether the dorsal claustrum receives afferent input from the intralaminar thalamic nuclei - centromedian nucleus, central lateral nucleus and paracentral nucleus. The intralaminar thalamic nuclei of eight cats were electrolytically lesioned. We obtained samples from the dorsal claustrum for electron microscopic analysis from the second to the seventh post-procedural day. Two types of degenerated synaptic boutons were observed: electron-dense which formed the majority of boutons, and electron-lucent comprising the remaining samples. Between the second and seventh post-procedural day, we observed a steady increase in the number of electron-dense boutons which were diffusely distributed throughout the dorsal claustrum. Electron-dense degenerated boutons formed asymmetrical contacts with dendritic spines as well as with small and medium-sized dendrites. In contrast, electron-lucent degenerated boutons were observed in earlier post-procedural periods and formed symmetrical axodendritic contacts.</p>","PeriodicalId":15966,"journal":{"name":"Journal of Histotechnology","volume":" ","pages":"117-125"},"PeriodicalIF":0.6,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140335942","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}
Elizabeth Chlipala, Tim Morken, Clare Thornton, Luis Chiriboga
{"title":"What will it take for histologists to be recognized under CLIA?","authors":"Elizabeth Chlipala, Tim Morken, Clare Thornton, Luis Chiriboga","doi":"10.1080/01478885.2024.2391636","DOIUrl":"10.1080/01478885.2024.2391636","url":null,"abstract":"","PeriodicalId":15966,"journal":{"name":"Journal of Histotechnology","volume":"47 3","pages":"93-96"},"PeriodicalIF":0.6,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142108085","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}