Journal of Histochemistry & Cytochemistry最新文献

{"title":"The Human Mechanosensory Corpuscles: A New Schwann Cell Localization of the Wilms' Tumor Protein WT1.","authors":"Alfonso Cepeda-Emiliani, María Otero-Alén, Tomás García-Caballero, Rosalía Gallego, Lucía García-Caballero","doi":"10.1369/00221554251338066","DOIUrl":"10.1369/00221554251338066","url":null,"abstract":"<p><p>SummaryThe Wilms' Tumor protein WT1 is a zinc-finger transcription factor with crucial roles in organogenesis, cell differentiation, tissue homeostasis, and oncogenesis. While its expression has been extensively studied in various tissues, its presence in the nervous system, particularly in peripheral glial cells, remains largely unexplored. In this study, we examined WT1 expression in the Schwann cells of mechanosensory corpuscles, nerve bundles, and free nerve endings (FNEs) within human penile tissues. Using single and double immunohistology, we analyzed WT1 coexpression with Schwann cell markers (S100, nestin, SOX10) and its association with axonal (neurofilaments, neuron-specific enolase, tyrosine hydroxylase) and perineurial/endoneurial markers (Glut-1, α-SMA, CD34). We found consistent WT1 cytoplasmic expression in the Schwann cells of Pacinian, Meissner, Krause, genital, Golgi-Mazzoni, and Ruffini-like corpuscles, with variable staining intensity. Confocal microscopy revealed WT1 colocalized with nestin but not S100, suggesting involvement in cytoskeletal organization. In addition, we documented WT1 in myelinating Schwann cells of nerve bundles, with distinct staining patterns in Cajal bands and Schmidt-Lanterman incisures, as well as in non-myelinating Schwann cells of FNEs. This is the first study to describe WT1 expression in sensory corpuscles, implicating it in Schwann cell development, maintenance, or plasticity, with potential relevance for peripheral nerve biology, pathology, and mechanosensation.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":" ","pages":"221554251338066"},"PeriodicalIF":1.9,"publicationDate":"2025-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12092412/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144110703","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparison of Manual Versus QuPath Software-based Immunohistochemical Scoring Using Oral Squamous Cell Carcinoma as a Model.","authors":"Hannah Horbas, Marcus Bauer, Alexander Eckert, Daniel Bethmann, Andreas Wilfer, Barbara Seliger, Claudia Wickenhauser","doi":"10.1369/00221554251335698","DOIUrl":"10.1369/00221554251335698","url":null,"abstract":"<p><p>Gold standard for immunohistochemical analyses is the manual assessment by two specialist pathologists. This process is time-consuming, highly dependent on the respective evaluator and often difficult to reproduce. The use of image analysis software, such as ImageJ, QuPath, or CellProfiler, which employ machine learning and/or deep learning mechanisms to perform biomarker analyses, offers a potential solution to these problems. The objective of our study is to evaluate whether digital assessment using the open-source software QuPath is comparable to manual evaluation and to examine the inter-evaluator variability between the two manual evaluators and two software-based evaluations. Six tissue microarrays (TMAs) were constructed for a cohort of 309 patients with primary oral squamous cell carcinoma (OSCC). The tumor tissue and corresponding non-lesional squamous epithelial mucosa specimen were immunohistochemically stained for the biomarkers Ki67, as a nuclear marker; the epidermal growth factor receptor (EGF-R), as a membranous marker; and the major histocompatibility complex class I (MHC-I) heavy chain (HC) expressed on the membrane and in the cytoplasm. The staining pattern was analyzed by two experienced, independent manual evaluators and by QuPath. The percentage of positive cells, for Ki67, and the histoscore (H-score) based on the percentage of positive cells and their staining intensity, for EGF-R and MHC-I, were determined as final values. The results yielded high to excellent spearman correlation coefficients for all three biomarkers (<i>p</i><0.001) in lesional and non-lesional tissues. The Bland-Altman plots demonstrated a high degree of agreement between manual and software-based analysis, as well as inter-evaluator variability demonstrating a high comparability of the evaluation methods. However, a prerequisite for a proper software-based analysis is an accurate, time-consuming annotation of the single specimen, which requires users with a comprehensive understanding of histology and extensive training in QuPath. Once these requirements are met, the software-based analysis offers advantages for large-scale biomarker studies due to objective and reproducible comparability of the stainings leading to a greater accuracy as well as the reuse of established conditions across similar analyses without requiring further operator input.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":" ","pages":"221554251335698"},"PeriodicalIF":1.9,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12081382/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144078211","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Expression of Mitochondrial Uncoupling Proteins and GABA Signaling Molecules in Unstimulated and Nerve Growth Factor-Stimulated PC12 Cells: Models for Chromaffin Cells and Sympathetic Neurons.","authors":"Keita Harada, Hidetada Matsuoka, Masumi Inoue","doi":"10.1369/00221554251332981","DOIUrl":"https://doi.org/10.1369/00221554251332981","url":null,"abstract":"<p><p>PC12 cells are a cell line originating from rat adrenal medullary chromaffin (AMC) cells. They extend a neurite-like structure in response to nerve growth factor (NGF). Thus, unstimulated and NGF-stimulated PC12 cells are used as models for AMC cells and sympathetic ganglion cells, respectively. However, how closely unstimulated and stimulated PC12 cells resemble AMC cells and sympathetic neurons, respectively, has not been elucidated sufficiently. We explored these issues by using biochemical and immunocytochemical methods. AMC cells and PC12 cells selectively expressed uncoupling protein 3 (UCP3) and uncoupling protein 4 (UCP4), respectively, and glucocorticoid activity inhibited UCP4 expression in PC12 cells. PC12 cells expressed extremely low levels of chromaffin granule-associated proteins, whereas the amount of synaptophysin, a synaptic vesicle-associated protein, was much higher than that in the adrenal medulla. Similar to AMC cells, the muscarinic receptor type 1 was located at the cell periphery in unstimulated PC12 cells, and its expression was markedly enhanced by NGF. Furthermore, NGF stimulation abolished the expression of GABA signaling molecules in PC12 cells. The results suggest that the properties of unstimulated PC12 cells are between those of AMC cells and sympathetic ganglion cells and GABA signaling is intrinsic to AMC cells.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":" ","pages":"221554251332981"},"PeriodicalIF":1.9,"publicationDate":"2025-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12037542/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143995776","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of Immunostaining Protocols for 3D Visualization of Pericytes in Human Retinal Flatmounts.","authors":"Noëlle Bakker, Aïcha A Croes, Eva Prevaes, Cornelis J F van Noorden, Reinier O Schlingemann, Ingeborg Klaassen","doi":"10.1369/00221554251323655","DOIUrl":"10.1369/00221554251323655","url":null,"abstract":"<p><p>Vascular pericytes are widely present across the human body and crucial in regulating vascular flow, permeability, and homeostasis. In the human retina, pericytes are important for forming and maintaining the blood-retinal barrier, as well as for autoregulation of blood flow. Pericyte loss has been implicated in various pathological conditions. Visualization of pericytes by immunofluorescence (IF) staining provides valuable information on pericyte number, morphology, location, and on expression of anatomic and functional markers. However, species-specific differences in pericyte marker expression exist. In this study, we aimed to develop a novel IF co-staining protocol to detect the pericyte markers NG2, PDGFRβ, αSMA, CD13, and RFC1 in human retinal flatmounts. Unlike retinal sections, retinal flatmounts enable 3D visualization of pericyte distribution across the entire vascular network. Key optimizations included tailoring the fixation method, blocking buffer composition and antibody solvent, as well as using jasplakinolide to enhance αSMA detection. Our protocol successfully enabled double staining of NG2 and PDGFRβ, as well as αSMA and PDGFRβ, whereas CD13 and RFC1 expression was not detectable in human retinal flatmounts. This novel 3D IF protocol enhances in situ visualization of human retinal pericytes, enabling accurate studies of their role in vascular health and disease to aid targeted therapy development.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":" ","pages":"147-170"},"PeriodicalIF":1.9,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11915233/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143649174","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Localization, Proteolytic Processing, and Binding Partners of Versican Isoforms in Vascular Lesions of Pulmonary Arterial Hypertension.","authors":"Christian Westöö, Ayse Ceren Mutgan, Oscar van der Have, Timothy J Mead, Salaheldin Ahmed, Elna Lampei, Christopher D Koch, Christian Norvik, Anders Aspberg, Martin Bech, Niccolò Peruzzi, Hans Brunnström, Grazyna Kwapiszewska, Göran Rådegran, Suneel S Apte, Karin Tran-Lundmark","doi":"10.1369/00221554251331271","DOIUrl":"https://doi.org/10.1369/00221554251331271","url":null,"abstract":"<p><p>Pulmonary arterial hypertension (PAH) is a lethal condition where expansion of the vascular extracellular matrix contributes to increased pulmonary vascular resistance. Versican, a chondroitin sulfate proteoglycan, is known to accumulate in vascular lesions of PAH and hyaluronan and tenascin-C, binding partners of versican, are elevated in PAH. The specific distribution and localization of versican isoforms, their cleavage products, and binding partners in vascular lesions of PAH had not been studied previously. Versican has five distinct isoforms, V0-V4, identified by the arrangement of its chondroitin-sulfate attachment regions, GAGα and GAGβ. Here, tissue from idiopathic PAH was imaged with synchrotron-based phase-contrast micro-CT and analyzed by histology, immunohistochemistry, and in situ hybridization. Plasma concentration of versican in PAH patients and controls was measured using ELISA. GAGα- and GAGβ-containing isoforms were identified in pulmonary arteriopathy of all patients. However, immunohistochemical staining of N-terminal G1 domain (versican G1) and C-terminal G3 domain (versican G3) using specific antibodies did not consistently co-localize. Tenascin-C was occasionally found in neointima, but also in thin-walled collateral vessels. Hyaluronan accumulated in the neointima, co-localizing with both versican G3 and the neoepitope DPEAAE. DPEAAE did not co-localize with the corresponding neoepitope of the C-terminal fragment generated by cleavage, possibly indicating motility of fragments. Patient plasma had a higher concentration of versican G3-containing fragments, compared to controls. The distribution of versican isoforms, cleavage products, and binding partners demonstrated here warrants further investigation of their functional roles in PAH, versican G3 was reinforced as a potential biomarker for PAH.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":"73 3-4","pages":"129-145"},"PeriodicalIF":1.9,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11993537/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144011272","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"MATLAB-based Methods Allow Precise, High-Throughput Quantification of Nuclear Morphology and Texture in Tachycardiomyopathy.","authors":"Moritz N S Mayer, Lisa M Köhler, Michael Paulus, Sabine Iberl, Maria Heinrich, Stefan Wagner, Lars S Maier, Alexander Dietl","doi":"10.1369/00221554251332331","DOIUrl":"https://doi.org/10.1369/00221554251332331","url":null,"abstract":"<p><p>The understanding of cardiomyopathies is hindered by a lack of quantitative histologic data. To address this methodical gap, we wrote a MATLAB-based image analysis platform to quantify nuclear and cellular disarray. We validated its utility in an animal model of tachycardiomyopathy (T-CM), whose ultrastructural remodeling processes have only partially been characterized and differ substantially from more prevalent cardiomyopathies. Six rabbits received right ventricular pacemaker implants. Three animals were paced incrementally up to 380 bpm for 30 days to induce T-CM. In three control rabbits, the pacemaker remained inactive (SHAM). Left ventricular tissue was collected, fixed in formalin, embedded in paraffin, stained, and digitized for nuclear morphometry, texture analysis, orientation analysis, and vascular architecture evaluation. Nuclear segmentation performed by the software was highly accurate, closely matching manual counts (mean manual nuclear count per slide = 81.3 ± 3.8, mean automated nuclear count per slide = 81.9 ± 4.3, <i>r</i> = 0.981, <i>p</i><0.001). In T-CM, nuclei were enlarged [SHAM (a.u.) = 2362, T-CM (a.u.) = 2660, <i>p</i>=0.0042]. Texture patterns differed between the groups with higher nuclear contrast in T-CM [SHAM (a.u.) = 0.0169, T-CM (a.u.) = 0.0247, <i>p</i>=0.0149], highlighting structural remodeling at the nuclear level. Median vessel size increased in T-CM [SHAM (a.u.) = 1532, T-CM (a.u.) = 2421, <i>p</i><0.0001]. In conclusion, our MATLAB-based image analysis platform allows high-throughput quantification of nuclear and extracellular disarray. It identified enlargement of nuclei and increased nuclear contrast as part of ultrastructural remodeling in tachycardiomyopathy.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":"73 3-4","pages":"115-128"},"PeriodicalIF":1.9,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11993534/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144008626","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Satellite DNA Amplification in Advanced Prostate Cancer Is Largely Independent From Euchromatic and Oncogene Amplicons.","authors":"Anja Weise, Antonio Augusto Ornellas, Gilda Alves, Constanze Pentzold, Jenny Holler, Melanie Wolter, Elena Jamali, Bernhard Theis, Thomas Liehr","doi":"10.1369/00221554251323657","DOIUrl":"10.1369/00221554251323657","url":null,"abstract":"<p><p>Recently, we were able to show that satellite DNA amplification (satDNA-AMP) is present in advanced prostate cancer. A chromosome microarray study provided first evidence that satDNA-AMP appears to be largely independent of centromere-near/pericentric euchromatic copy number alterations. Therefore, it might be carefully suggested that satDNA-AMP could be a new and independent marker for advanced tumor progression.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":" ","pages":"109-113"},"PeriodicalIF":1.9,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11915227/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143649186","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Histo-LOOP: A Novel Embedding Tool for Standardizing, Simplifying, and Advancing Histological Tissue Preparation.","authors":"Pakhwan Nilcham, Mamdouh Afify, Nicole Schaaps, Carolina Neu, Rahma Shahin, Andreas Prescher, Felix Jan Vogt, Anne Turoni-Glitz","doi":"10.1369/00221554251329978","DOIUrl":"10.1369/00221554251329978","url":null,"abstract":"<p><p>Tissue preparation for paraffin embedding is a crucial step in histological processes. Standardized methods are required to ensure the accuracy of research and clinical diagnostic results. However, standardization is particularly challenging for long luminal tissues. Conventional methods such as single/serial sections and the Swiss roll, often have drawbacks including the risk of missing or misaligned sections, excess consumption of materials, and high workload. They also require significant expertise and are difficult to standardize. To address these issues, we developed the Histo-LOOP embedding tool-a novel tool designed to standardize, simplify, and improve histological processing. Histo-LOOP is suitable for various tissue types including long tubular tissue, allowing for a complete overview in cross-sectional and longitudinal views. It is also suitable for punch biopsies or small sections, and enables the assessment of multiple punch biopsies or sections within one paraffin block, and in multiple cutting planes, for example, for liver and prostate biopsies. Histo-LOOP does not interfere with the sectioning and staining process and does not cause artifacts.Here, we introduce the novel tool Histo-LOOP and describe preparation techniques for tubular tissue and small tissue samples using this tool, along with examples of their histological evaluation.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":" ","pages":"97-107"},"PeriodicalIF":1.9,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11955986/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143742897","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dominant Expression of Chromogranin B in Pituitary Corticotrophs and Its Putative Role in Interaction With Secretogranin III.","authors":"Shota Kikuchi, Koki Odashima, Tadashi Yasui, Seiji Torii, Masahiro Hosaka, Hiroshi Gomi","doi":"10.1369/00221554241311965","DOIUrl":"10.1369/00221554241311965","url":null,"abstract":"<p><p>SummaryPrevious studies have suggested that chromogranin A (CgA) is a partner molecule of secretogranin III (SgIII). In mouse pituitary corticotroph-derived AtT-20 cells, SgIII plays a role in sorting CgA/hormone aggregates into secretory granules (SGs). Although CgA expression is equivocal, CgB is clearly detectable in the rat pituitary corticotrophs. Therefore, we hypothesized that CgB shares a function with CgA in pituitary corticotrophs. In the binding assays, CgB, similar to CgA, showed binding activity to SgIII under weakly acidic conditions and in the presence of Ca²⁺. Considering the differences in animal species, the different abilities of antibodies, and the conditions of tissue fixation and thin sectioning in immunofluorescence histochemistry, we found that CgA was expressed in a small population (approximately 10%), and its expression intensity was weaker than that of CgB (>98%) in rodent pituitary corticotrophs. In addition, similar to CgA, CgB and SgIII were colocalized in adrenocorticotropic hormone (ACTH) granules. The labeling of CgA and CgB was not completely consistent, and CgB colocalized with SgIII in many granules. These results suggest that there are multiple sorting systems for ACTH granules in pituitary corticotrophs and that the SgIII/CgB complex behaves more dominantly than the SgIII/CgA complex, which has somewhat different properties.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":" ","pages":"29-53"},"PeriodicalIF":1.9,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11719422/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142950146","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"RNAscope Multiplex FISH Signal Assessment in FFPE and Fresh Frozen Tissues: The Effect of Archival Duration on RNA Expression.","authors":"Ariestya Indah Permata Sari, Katherine Copeland, Pattarin Nuwongsri, Wiriya Pipatsakulroj, Artit Jinawath, Nipan Israsena, Panuwat Lertsittichai, Prakasit Chirappapha, Meng-Shin Shiao, Natini Jinawath","doi":"10.1369/00221554241311971","DOIUrl":"10.1369/00221554241311971","url":null,"abstract":"<p><p>Formalin-fixed paraffin-embedded tissue (FFPET), which is the most widely used pathology archive, usually has low-quality DNA and RNA due to extensive nucleic acid crosslinking. RNA fluorescence in situ hybridization (RNA-FISH) has been increasingly utilized in research and clinical settings to diagnose disease pathology. In this study, the effect of RNA degradation over archival time on RNA-FISH signals in FFPET and fresh frozen tissue (FFT) was systematically assessed. RNAscope multiplex fluorescent assay with the four house-keeping-gene (HKG) probes <i>UBC, PPIB, POLR2A</i>, and <i>HPRT1</i> was performed on 62 archived breast cancer samples (30 FFPETs and 32 FFTs). As expected, the number of RNAscope signals in FFPETs is lower than in FFTs in an archival duration-dependent fashion. The RNA degradation in FFPETs is most pronounced in high-expressor HKGs, <i>UBC</i> and <i>PPIB</i>, than in low-to-moderate expressors <i>POLR2A</i> and <i>HPRT1</i> (<i>p</i><0.0001). Analysis of RNA expression over time showed that <i>PPIB</i>, which has the highest signal, was the most degraded in both adjusted transcript and H-score quantification methods (<i>R</i>² = 0.35 and <i>R</i>² = 0.33, respectively). This proves that although the RNAscope probes are designed to detect fragmented RNA, performing a sample quality check using HKGs is strongly recommended to ensure accurate results.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":" ","pages":"9-28"},"PeriodicalIF":1.9,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11755420/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143023699","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}