Journal of Histochemistry & Cytochemistry最新文献

{"title":"Phosphotungstic Acid-treated Picrosirius Red Staining Improves Whole-slide Quantitative Analysis of Collagen in Histological Specimens.","authors":"Yui Mukade, Sayaka Kobayashi, Yoshimi Nishijima, Kiminori Kimura, Akira Watanabe, Hayato Ikota, Ken Shirabe, Hideaki Yokoo, Masanao Saio","doi":"10.1369/00221554221141140","DOIUrl":"10.1369/00221554221141140","url":null,"abstract":"<p><p>We tried to prevent nonspecific nuclear staining (NS-NS) of picrosirius red (PSR) staining by treating the specimens with one of the heteropoly acids phosphotungstic acid (PTA). We analyzed a total of 35 cases of non-cancerous liver tissue for fibrosis and NS-NS under PSR-alone, phosphomolybdic acid (PMA)-pretreated PSR (PMA + PSR), or PTA-pretreated PSR (PTA + PSR) condition. In addition, we analyzed the photosensitivity of PMA or PTA single stain specimens. PTA + PSR significantly suppressed NS-NS compared with PSR. The color of the specimens did not change into blue by 30 times the exposure to whole slide scanner (WSS) light. The PTA + PSR condition showed the highest correlation with the Ishak score (pathological evaluation of liver fibrosis) compared with other conditions. Furthermore, Sirius Red-positive percentage (SRP%) in PSR was increased in the NS-NS observed cases. SRP% in PMA + PSR was significantly affected by WSS light exposure time. Moreover, the deposition of non-polarized PSR-stained substances (NP-PSR⁺S) clinging to the collagen fibers potentially explains why SRP% seemed bigger under PSR than PTA + PSR. Our protocol enabled us to analyze the whole slide image of PSR staining by high magnification, which would contribute to the accurate analysis of collagen amount in the tissue sections.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9912349/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10713178","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":"Commentary on \"Classifications of Anterior Pituitary Cell Types With Immunoenzyme Histochemistry\": Dr. Paul Nakane Blazed the Trail to Modern Technology.","authors":"Gwen V Childs","doi":"10.1369/00221554221146837","DOIUrl":"10.1369/00221554221146837","url":null,"abstract":"","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9912347/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9858054","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":"The Graham and Karnovsky Horseradish Peroxidase Ultrastructural Method: A Premier JHC Citation Classic.","authors":"Dale R Abrahamson","doi":"10.1369/00221554221146838","DOIUrl":"10.1369/00221554221146838","url":null,"abstract":"<p><p>This commentary briefly reviews the background for the development of the horseradish peroxidase-diaminobenzidine tetrahydrochloride histochemical method originally described by Graham and Karnovsky in their citation classic, reprinted in full in this issue of <i>Journal of Histochemistry & Cytochemistry</i>. Some of the method's subsequent applications, including its use as a macromolecular tracer for kidney glomerular permeability and use in immunoelectron microscopy and other immunoassays, are also discussed.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9912348/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10710504","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":"A Forgotten Principle in Immunocytochemistry: Optimal Dilution.","authors":"Bertalan Dudás,&nbsp;Malcolm Lane,&nbsp;Nikitha Mupparaju,&nbsp;Hye Min Kim,&nbsp;Istvan Merchenthaler","doi":"10.1369/00221554221146213","DOIUrl":"10.1369/00221554221146213","url":null,"abstract":"<p><p>Immunocytochemical (ICC) techniques are frequently used in basic and clinical research. Here, we focus on the importance of using antisera/antibodies at optimal dilutions to achieve specificity and reduce costs. Unfortunately, the basic principle, the necessity to test method specificity of the staining by a series of increasing dilutions of primary antiserum/antibodies, is only occasionally seen in papers using ICC. Many researchers rely on the company's information or others' published data. In this study, we show examples with monoclonal antibodies used in the peroxidase-based ICC technique in mouse and guinea pig brain sections. We show images of ICC staining of phospho-S129 alpha-synuclein in A53T mice and NeuN in guinea pig brains and demonstrate that optimal staining with them can be achieved at least at two to three orders of magnitude higher dilutions than generally used in the literature. We strongly recommend that when antisera/antibodies are used for the first time in any laboratory, independent of what the manufacturer or vendor recommends or are found in the literature, a dilution curve should be set up to identify the optimal dilution. This practice provides not only the highest specificity but is also an economic approach.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2022-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9903208/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10713953","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":"Commentary on \"Antigen Retrieval Immunohistochemistry: Past, Present, and Future\".","authors":"Clive R Taylor,&nbsp;Shan-Rong Shi","doi":"10.1369/00221554221147086","DOIUrl":"10.1369/00221554221147086","url":null,"abstract":"Almost half a century has passed from the first successful application of peroxidase labeled antibodies to routinely processed formalin-fixed paraffin-embedded (FFPE) tissues with the express purpose of improved diagnosis. Retention of the morphological features that are the foundation of surgical pathology was key to rendering the “immunoperoxidase” method attractive for diagnostic use. However, significant obstacles remained, including the limited number of antibodies that “worked” on FFPE tissues, the relatively low sensitivity of detection methods, and, most important in terms of broad utility, the deleterious effects of formalin fixation on many antigens. There were advances, some dramatic, but overall progress was slow.1 The advent of the hybridoma technique was eventually to lead to the generation of numerous monoclonal antibodies that were rapidly incorporated into immunohistochemistry. Concurrently, detection systems improved, and attempts were made to “undo” the effects of formalin fixation by enzymatic digestion, with limited success. Against this background, the introduction of what now is known as “antigen retrieval” (AR), or by some as heat induced epitope retrieval (HIER), provided huge impetus to the field, having the effect of greatly increasing the number of antigens that were demonstrable in FFPE tissues, including many with diagnostic application. This advance was entirely counterintuitive and was based upon exhaustive research in dusty libraries (no Internet searches!). There evidence was found that the “antigenic potency” of tetanus toxoid, toxin that had been inactivated by formalin, could in large part be restored by something as simple as boiling. Why not apply the same approach to FFPE tissues? The current issue of the Journal of Histochemistry & Cytochemistry has selected as its Classical Article a paper by Shan-Rong Shi, Richard Cote, and Clive Taylor that describes the evolution of the AR method, and the impact of AR in diagnostic pathology and broader fields of research, as measured just 5 years after its introduction. That this initial promise has been fulfilled manyfold is attested by extensive day-to-day use of AR in histochemistry laboratories worldwide, and by an enormous ever expanding literature, even providing the basis for DNA extraction methods that 1147086 JHCXXX10.1369/00221554221147086Antigen Retrieval: A Simple Idea With Major Ongoing ImpactTaylor and Shi article-commentary2022","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2022-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9903211/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10714434","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":"Commentary on \"Application of Photoshop-based Image Analysis to Quantification of Hormone Receptor Expression in Breast Cancer\".","authors":"Allen M Gown","doi":"10.1369/00221554221145227","DOIUrl":"10.1369/00221554221145227","url":null,"abstract":"At the time of publication of this article, image analysis as applied to diagnostic surgical pathology was a nascent technology, often characterized as a “solution in search of a problem.” After all, biomarkers such as estrogen receptor (ER) in breast cancer were scored in a threshold manner, with the assumption that this scoring was highly accurate and reproducible. Unfortunately, several studies in the early 2000s found significant discordance in this ER evaluation when slides were re-reviewed.1,2 Ironically, prior approaches to determining ER expression in breast cancer, for example, the dextran-coated charcoal method, were quantifiable and reproducible, but required fresh frozen tissue and could not be used on formalin-fixed paraffin-embedded tissue. Image analysis on deparaffinized, formalin-fixed tissue sections immunostained with appropriate antibodies was anticipated to be a method that could increase the predictive power of IHC determination of hormone receptor status. Indeed, this 1997 paper was one of the first to employ image analysis in the evaluation of ER. While after more than two decades Photoshop has maintained its role as a premier raster graphics and digital art editor, it has not taken a second life as an image analysis tool. We selected Photoshop as a “maverick” technique that permitted us to accomplish the analysis we desired, but to also avoid the use of commercial image analysis instruments and software, which at that time were expensive and unaffordable for our laboratory. Today, while commercial image analysis hardware and software are legion, free public domain software such as QuPath can yield comparable results as many commercial software packages3 and far exceed the capabilities of Photoshop. While our 1997 paper succeeded as a “proof of principle,” now 25 years later, ER is still not quantified in the vast majority of cases, but is instead still “eyeballed” to determine whether more than 1% of tumor cells are positive for nuclear signal (or between 1% and 10%), but no image analysis–based quantification is required.4 There may still be a role in the future for more quantitative ER determination in breast cancer, but that will be determined not by technology alone but largely by evidence that may be forthcoming from clinical studies. But it will almost certainly not involve the use of Photoshop!","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2022-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9903209/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9280808","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":"Long Hanging Structure of Collagen VII Connects the Elastic Fibers and the Basement Membrane in Young Skin Tissue.","authors":"Takeshi Tohgasaki,&nbsp;Shino Nishizawa,&nbsp;Shinya Kondo,&nbsp;Shioji Ishiwatari,&nbsp;Tetsuhito Sakurai","doi":"10.1369/00221554221145998","DOIUrl":"10.1369/00221554221145998","url":null,"abstract":"<p><p>Aging leads to substantial structural changes in the skin. Elastic fibers maintain skin structure, but their degeneration and loss of function with age result in wrinkle formation and loss of skin elasticity. Oxytalan fiber, a type of elastic fiber, extends close to the dermal-epidermal junction (DEJ) from the back of the dermis. Oxytalan fibers are abundant in the papillary layer and contribute to skin elasticity and texture. However, to accurately understand the mechanisms of skin elasticity, the interaction between elastic fibers and DEJ should be elucidated. Here, we investigated elastic fibers and DEJ and their structural alterations with aging. Several basement membrane proteins [collagen (COL) IV, COLVII, and laminin 332], fibrous tropoelastin, and fibrillin-1 in excised human skin tissue were observed using three-dimensional imaging. Age-related alterations in COLVII, elastic fibers, and fibrillin-1 were evaluated. We found that COLVII forms long hanging structures and is co-localized with fibrous tropoelastin in young skin but not aged skin. Fibrillin-1-rich regions were observed at the tips of elastin fibers in young skin tissue, but rarely in aged skin. This co-localization of elastic fiber and COLVII may maintain skin structure, thereby preventing wrinkling and sagging. COLVII is a potential therapeutic target for skin wrinkling.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2022-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9903210/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10710505","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":"Abnormal Expression of an Insulin Synthesizing Enzyme in Islets of Adult Autoantibody Positive Donors.","authors":"Gladys Teitelman","doi":"10.1369/00221554221138368","DOIUrl":"10.1369/00221554221138368","url":null,"abstract":"<p><p>The observation that the two active forms of proprotein convertase 1/3 (PC1/3) were differentially expressed in beta cells of normal islets raised the possibility that this heterogeneity is lost during type 1 diabetes (T1D) progression. To test this hypothesis, the expression of the convertase was evaluated by confocal microscopy in sections of human pancreas of autoantibody positive (AA+) and T1D donors and compared with that of control. Islets of T1D pancreas were comprised of beta cells expressing either low or high PC1/3 levels and all islets of a pancreatic section contained only one beta cell type. Pancreata of AA+ donors contained either of these two classes of islets intermixed with normal islets comprised of beta cells with heterogeneous PC1/3 expression. This alteration affected the expression of proinsulin and insulin, which in most AA+ and T1D donors were lower than in controls. The present results indicate that the heterogeneity of PC1/3 expression is lost in all beta cells in a subset islets of AA+ donors and in all islets of T1D donors. These findings suggest that the heterogeneity of PC1/3 expression is a biomarker of human beta cell health and that its loss coincides with the initial stages of T1D.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2022-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9660365/pdf/10.1369_00221554221138368.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9493247","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":"Aberrant Expression of Thymosin Beta-4 Correlates With Advanced Disease and BRAF V600E Mutation in Thyroid Cancer.","authors":"Chi-Yu Kuo,&nbsp;Jie-Yang Jhuang,&nbsp;Wen-Chien Huang,&nbsp;Shih-Ping Cheng","doi":"10.1369/00221554221138370","DOIUrl":"10.1369/00221554221138370","url":null,"abstract":"<p><p>Thymosin beta-4 (TMSB4X) was recently identified as a differentially expressed gene between malignant and non-malignant thyroid cells via single-cell RNA sequencing. In the present study, we aimed to study the immunostaining pattern of TMSB4X in benign and malignant thyroid neoplasms. Immunohistochemical analysis revealed that normal thyroid tissue or benign thyroid disorders exhibited undetectable immunoreactivity against TMSB4X except for positive staining of inflammatory infiltrates and stromal cells associated with autoimmune thyroid disease. By contrast, overexpression of TMSB4X was observed in a variety of thyroid malignancies, including papillary, follicular, poorly differentiated, and undifferentiated thyroid cancer. Among 141 patients with differentiated thyroid cancer, higher TMSB4X expression was associated with papillary tumor type, extrathyroidal extension, lymph node metastasis, and BRAF V600E mutation. The results were consistent with those from the public transcriptomic datasets. In summary, TMSB4X expression was aberrantly increased in various types of thyroid cancer, and higher TMSB4X expression was correlated with advanced disease characteristics. Thymosin beta-4 may be a novel downstream effector of the BRAF V600E mutation.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2022-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9660367/pdf/10.1369_00221554221138370.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10635856","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 and Clinical Significance of Spi-B in B-cell Acute Lymphoblastic Leukemia.","authors":"Yuzuru Ariga,&nbsp;Shulin Low,&nbsp;Hitomi Hoshino,&nbsp;Tsutomu Nakada,&nbsp;Tomoya O Akama,&nbsp;Akifumi Muramoto,&nbsp;Mana Fukushima,&nbsp;Takahiro Yamauchi,&nbsp;Yusei Ohshima,&nbsp;Motohiro Kobayashi","doi":"10.1369/00221554221130383","DOIUrl":"10.1369/00221554221130383","url":null,"abstract":"<p><p>Spi-B, a member of the E26 transformation-specific (ETS) family of transcription factors, plays an important role in B cell differentiation. Spi-B also functions in development of diffuse large B-cell lymphoma; thus, we hypothesized that it may participate in leukemogenesis of B-cell acute lymphoblastic leukemia (B-ALL). To test this hypothesis, we first generated an anti-Spi-B monoclonal antibody that recognized Spi-B on formalin-fixed, paraffin-embedded tissue sections. This antibody, designated S28-5, selectively stained B cell nuclei at the pre-plasma cell stage (including centrocytes and centroblasts in germinal centers) and nuclei of plasmacytoid dendritic cells, but not fully differentiated plasma cells, T cells, macrophages, or follicular dendritic cells. Employing S28-5, we then performed immunohistochemical staining of bone marrow aspiration biopsy specimens obtained from B-ALL patients (<i>n</i>=62). Cases that showed stronger nuclear S28-5 signals than T-cell ALL were scored positive. In 26 (42%) of 62 specimens, leukemic cells showed nuclear Spi-B expression, and positivity was associated with patient age at diagnosis, and serum uric acid and creatinine levels. Moreover, Spi-B-positive patients demonstrated significantly shorter overall survival than did Spi-B-negative patients. These results suggest that Spi-B expression may serve as a prognostic indicator of B-ALL.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2022-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9660366/pdf/10.1369_00221554221130383.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40379552","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}