Готові списки джерел за темами / Immunohistochemistry

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Добірка наукової літератури з теми "Immunohistochemistry"

Автор: Grafiati

Опубліковано: 4 червня 2021

Оновлено: 30 липня 2024

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Статті в журналах з теми "Immunohistochemistry"

B. Ingle, Sachin. "Immunohistochemistry." International Journal of Current Research and Review 10, no. 11 (2018): 0–1. http://dx.doi.org/10.31782/ijcrr.2018.10115.

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Taylor, Clive R. "Immunohistochemistry." Applied Immunohistochemistry & Molecular Morphology 27, no. 5 (2019): 325–26. http://dx.doi.org/10.1097/pai.0000000000000770.

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Mani, Haresh, and Dani S. Zander. "Immunohistochemistry." Chest 142, no. 5 (November 2012): 1324–33. http://dx.doi.org/10.1378/chest.12-0123.

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&NA;. "Immunohistochemistry." Pathology 22 (1990): 9–10. http://dx.doi.org/10.3109/00313029009060103.

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Aryal, G. "Immunohistochemistry." Journal of Pathology of Nepal 5, no. 10 (September 14, 2015): I. http://dx.doi.org/10.3126/jpn.v5i10.15660.

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Анотація:

Immunohistochemistry (IHC) or immunocytochemistry is a method of localizing specific antigen in tissue or cells based on antigen antibody reaction. IHC is the way of validating morphological findings. It helps in tumor diagnosis and classification, identify prognostic and predictive markers. IHC has a long history that dates back more than 70 years, when Coons1 first developed immunofluroscence technique to detect corresponding antigen in frozen tissue section. At oxford, Taylor and Burns2 developed the first successful demonstration of antigens in routinely processed formalin fixed paraffin-embedded sections. Since the early 1990s, IHC has been applied in routine formalin fixed paraffin embedded tissue. 3-4 Validation of reagents, protocols, controls and staining results are vital steps of IHC. The basic principles and protocols for fresh-frozen tissue sections are the same as those for paraffin sections, except that the antigen retrieval and dewaxing procedures are not required for frozen tissue sections. Titrations may also differ and must be separately optimized. Basic principles: Antigen-antibody recognition is based on the three-dimensional (3D) structure of protein or some other antigen, which may be compromised by formalin-induced modification of protein conformation (“masking”) but is restored in part by Antigen retrieval. Anti-A antibody binds specifically to antigen A in the tissue section. Antigen B (B) is depicted as a second antigenic determinant that is part of the anti-A molecule; anti-B antibody, made in a second species, will bind to this determinant. Thus anti-B, the so-called secondary antibody, can be used to locate the site of binding of anti-A, the primary antibody, in a tissue section. Basic IHC procedure Antigen retrieval (AR)-Enzymatic digestion (proteinase or trypsin), heat treatment (Microwave, water bath or autoclave) Blocking of non-specific background staining Incubation with primary antibody in humidity chamber Add avidin-biotin-peroxidase complex, which binds to secondary antibody Add 3, 3’ diaminobenzidine (DAB) as a chromagen (color changing reagent), with hematoxylin (Mayer's) counterstaining Theranostic Application IHC is becoming increasingly important for the evaluation of predictive markers that can help select patients who may respond to particular targeted therapies. Some of these CD117 for GI stromal tumors, Herceptin (Genentech, South San Francisco, CA) for HER2-positive breast cancers, and rituximab for CD20-positive lymphomas.

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Dabbs, David J. "Immunohistochemistry." Pathology Case Reviews 4, no. 6 (November 1999): 229. http://dx.doi.org/10.1097/00132583-199911000-00001.

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Mani, Haresh, and Dani S. Zander. "Immunohistochemistry." Chest 142, no. 5 (November 2012): 1316–23. http://dx.doi.org/10.1378/chest.11-3327.

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Mazroa, Shireen A. "Immunohistochemistry." Egyptian Journal of Histology 35, no. 2 (June 2012): 191–97. http://dx.doi.org/10.1097/01.ehx.0000414291.44156.ef.

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Wittekind, Christian, and Andrea Tannapfel. "Immunohistochemistry." Digestion 58, no. 1 (1997): 79–81. http://dx.doi.org/10.1159/000201534.

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Sompuram, Seshi R., Kodela Vani, Brian Tracey, Debra A. Kamstock, and Steven A. Bogen. "Standardizing Immunohistochemistry." Journal of Histochemistry & Cytochemistry 63, no. 9 (May 4, 2015): 681–90. http://dx.doi.org/10.1369/0022155415588109.

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Дисертації з теми "Immunohistochemistry"

King, George. "Studies of amplification methods in immunohistochemistry." Thesis, University of Aberdeen, 2001. http://digitool.abdn.ac.uk/R?func=search-advanced-go&find_code1=WSN&request1=AAIU537963.

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Анотація:

The aim of this project was to compare and contrast the results of three related methodologies which are of particular relevance or potential within our department -. a manually performed streptAvidin Biotin Complex (sABC/ HRP) method which for many years has served as our 'standard' immunocytochemical method; an integrated semi-automated systems from DAKO based on the TechMate 500 immunostainer and the ChemMate reagent package;. a manually performed tyramide amplification method regarded as amongst the most sensitive immunocytochemical method currently available. Antigen retrieval investigating differing durations of exposure to the proteolytic enzyme trypsin, and heat mediated antigen retrieval using citrate buffer pH6.0 or DAKO high pH antigen retrieval solution were also included as parameters for investigation. To this aim a panel of eleven specific antibodies were assessed, each of which had previously displayed individual qualities worthy of investigation -. PGP9.5, CD30, oestrogen receptor, CD3 polyclonal, CD2, CD3 monoclonal, CD4, cyclin D1, kappa immunoglobulin light chain, lambda immunoglobulin light chain and IgD immunoglobulin heavy chain. The results of this work demonstrate that each of the systems investigated has its individual merits -. The manual sABC/ HRP method is the least complicated and expensive of the methods to perform. There are significant disadvantages in its used however. Being a manual method there are many steps where individual variation in working practices within a laboratory can and do affect results. The TechMate/ ChemMate system is a more sensitive system than the manual sABC/ HRP method also providing a cleaner end result and allowing higher dilutions of primary antibody to be employed. Tyramide amplification is the most sensitive system evaluated, the antibodies in this work generally demonstrated immunostaining at higher dilutions than the other methods tested although some, in particular those directed against immunoglobulin light chains, operated better under the ChemMate/ TechMate system. On occasion specific positive results were obtained which none of the other methods tested could achieve with certain antibodies, CD3 monoclonal and to a lesser extent CD2.

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Lam, Sing-chi, and 藍承志. "A HRCT and immunohistochemistry study on bronchiectasis." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2004. http://hub.hku.hk/bib/B30402499.

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Shu, Jie. "Immunohistochemistry image analysis : protein, nuclei and gland." Thesis, University of Nottingham, 2014. http://eprints.nottingham.ac.uk/27616/.

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This thesis focus on the analysis of digitized microscopic image, especially on IHC stained colour images. The corresponding contributions focused on the automatic detection of stain colour and glands, the segmentation and quantification of cell nuclei, the analysis of liver cirrhosis and the development of a semi-automatic toolbox. Colour is the most important feature in the analysis of immunostained images. We developed a statistical colour detection model for stain colour detection based on the histograms of collected colour pixels. This is acting on the approach "what you see is what you get" which outperforms the other methods on the detection of several kinds of stain colour. Verifying the presence of nuclei and quantifying positive nuclei is the foundation of cancer grading. We developed a novel seeded nuclei segmentation method which greatly improves the segmentation accuracy and reduces both over-segmentation and under-segmentation. This method has been demonstrated to be robust and accurate in both segmentation and quantification against manual labelling and counting in the evaluation process. The analysis of gland architecture, which reflects the cancer stage, has evolved into an important aspect of cancer detection. A novel morphology-based approach has been developed to segment gland structures in H-DAB stained images. This method locates the gland by focusing on its morphology and intensity characteristics, which covers variations in stain colours in different IHC images. The evaluation results have demonstrated the improvements of accuracy and efficiency. For the successive development of three methods, we put them in a semi-automatic toolbox for the aid of IHC image analysis. It can detect different kinds of stain colour and the basic components in an IHC image. The user created models and parameters can be saved and transferred to different users for the reproduction of detection results in different laboratories. To demonstrate the flexibility of our developed stained colour detection technique, the tool has been extended to the analysis of liver cirrhosis. It is a novel method based on our statistical colour detection model which greatly improves the analysis accuracy and reduces the time cost.

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Koria, Muntaha. "Identification of PHPT1 in mouse tissues by immunohistochemistry." Thesis, Uppsala University, Department of Medical Biochemistry and Microbiology, 2007. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-7745.

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Анотація:

Although it has been estimated that protein histidine phosphorylation account for about 6 % of the protein phosphorylation in eukaryotic cells; the knowledge of histidine phosphorylation and dephosphorylation is still limited. Lately, studies have appeared of a mammalian 14-kDa phospho- histidine phosphatase, also named protein histidine phosphatase and molecular cloning have provided some information of its physiological role. The object of the present study was to detect the protein expression of protein histidine phosphatase, PHPT1, in mouse tissue, by using immunohistochemistry. Tissue samples from a 4-week-old mouse (heart, liver, kidney, lung, muscle, and spleen), 5-month-old mouse (testis and intestinal), 8-month-old mouse (uterus) and an embryo from 14.5 days old mouse were obtained and processed for light microscopic examination. An absorption test was also made to confirm the specificity of the antibody. The results reveal that PHPT1 is mainly expressed in epithelium, heart- and skeletal muscle. These results provide new evidences for the understanding of the function of eukaryotic histidine phosphorylation and dephosphorylation.

KEYWORDS

Phosphohistidine, dephosphorylation, protein histidine phosphatase, phosphohistidine phosphatase, protein phosphorylation

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Sundara, Rajan Sreekumar. "Investigating male breast cancer using transcriptomics and immunohistochemistry." Thesis, University of Leeds, 2016. http://etheses.whiterose.ac.uk/15847/.

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Background: The rare nature of male breast cancer (MBC) has led to its management being guided by the extensive research conducted in the field of female breast cancer (FBC). The aim of this study was to evaluate MBC at both protein and molecular level to improve understanding of its pathology. Methodology: Immunohistochemistry analysis was performed in MBC (n=428) TMAs for 18 biomarkers (ERα, ERβ1, ERβ2, ERβ5, Total PR, AR, CK5/6, CK14, CK18, CK19, p53, Bcl-2, Her2, E-cadherin, Ki67, Survivin, Prolactin and FOXA1). The manual scoring of ERα and Ki67 was correlated with a fully automated immunohistochemistry image analysis system (ImmunoRatio™). Finally gene expression profiling (GEP) was undertaken in matched MBC (n=15) and FBC (n=10) samples. Results: There was poor 5 year overall survival (OS) in CK18 and CK19 negative patients (p= 0.05; p= 0.003), as well as poor 10 year OS in CK19 negative patients (p= 0.002). Age (p= 0.001) and nodal status (p= 0.04) was found to be independent predictors of OS at 5 years. There was significant correlations between manual and ImmunoRatio™ ERα (ρ= 0.872; p= 0.000) and Ki67 (r= 0.675; p= 0.000) scores. However due to a low measure of agreement it was not possible to validate Ki67 scoring using ImmunoRatio™. The functional enrichment analysis of GEP data using less stringent criteria (p < 0.05) identified 735 differentially expressed genes. The data analysis showed up-regulation of genes involved in ECM synthesis, degradation and re-modelling in MBC. The end product of one of the up-regulated genes (Fibronectin (FN1)) was validated in the MBC cohort with high fibronectin expression (60%) being positively associated with nodal status and showed a trend towards poor 5 year OS (p= 0.06). Conclusion: In MBC, epithelial cytokeratins, especially CK19 was found to be of prognostic significance. The extracellular matrix remodelling associated genes were found to be up-regulated in MBC. Fibronectin, end product of one of the up-regulated gene was found to have prognostic significance in MBC.

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Café, Marçal Valéria. "Pathology and immunohistochemistry of Cheetah (Acinonyx jubatus) myelopathy /." [S.l.] : [s.n.], 2006. http://www.stub.ch/index.php?p=1&i=645.

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Rozell, Björn. "Immunohistochemical studies of the thioredoxin system." Göteborg : Dept. of Histology, University of Göteborg, 1987. http://catalog.hathitrust.org/api/volumes/oclc/17242526.html.

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Kolivras, Athanassios. "Immunohistochemistry in the histopathological diagnosis of primary scalp alopecia." Doctoral thesis, Universite Libre de Bruxelles, 2016. http://hdl.handle.net/2013/ULB-DIPOT:oai:dipot.ulb.ac.be:2013/238160.

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Primary scalp alopecia is classically divided into cicatricial (scarring) and non-cicatricial (non-scarring). Challenging cases are assessed with a scalp biopsy. The use of both horizontal and vertical sections (HoVert sections) has dramatically improved the accuracy of histopathological diagnosis. In this work, we have used immunostaining to address diagnostic difficulties, which persist despite all currently available tools. We performed an immunostain panel (CD3, CD4, CD8 and CD20) in order to distinguish pattern hair loss from alopecia aerate in cases which do not have the usual peribulbar lymphocytic infiltrate and showed that CD3+ T-lymphocytes within the empty fibrous follicular tracts favor a diagnosis of alopecia areata. We performed CD123 in order to distinguish lichen planopilaris from alopecia lupus erythematosus in cases with only a superficial lymphocytic infiltrate and an uninvolved interfollicular epidermis and showed that clusters of CD123+ plasmacytoid dendritic cells favor a diagnosis of lupus erythematosus. We performed cytokeratin 15 in order to assess whether the loss of the follicular bulge stem cells has diagnostic value in cicatricial alopecia and demonstrated that the loss of cytokeratin 15+ bulge stem cells is identified in lichen planopilaris, frontal fibrosing alopecia, and lupus erythematous, so cytokeratin 15 has no diagnostic value. We have attempted to integrate the new concepts and our findings into the traditional classifications of alopecia and proposed a new diagnostic algorithm. In conclusion, immunostaining combined with HoVert grossing advances the accuracy of histopathological diagnosis of primary scalp alopecia.
L’alopécie primitive du cuir chevelu est habituellement classée en cicatricielle et non-cicatricielle. Dans les cas difficiles, la biopsie du cuir chevelu peut aider au diagnostic. L’utilisation de coupes, à la fois verticales et horizontales sur le même spécimen (technique HoVert), a radicalement amélioré le diagnostic histopathologique. Dans ce travail, nous avons utilisé l’immunohistochimie pour évaluer les difficultés diagnostiques qui persistent malgré tous les outils actuels. Nous avons utilisé les CD3, CD4, CD8 et CD20 pour différencier l’alopécie androgénique de la pelade dépourvue de l’infiltrat lymphocytaire péribulbaire habituel et nous avons démontré que la présence de lymphocytes CD3+ dans les travées folliculaires fibreuses est en faveur de la pelade. Nous avons utilisé le CD123 pour différencier le lichen plan pilaire du lupus érythémateux alopécie avec infiltrat lymphocytaire superficiel et sans atteinte de l’épiderme interfolliculaire et nous avons démontré que la présence d’amas de cellules dendritiques plasmacytoïdes CD123+ est en faveur du lupus érythémateux. Nous avons utilisé la cytokératine 15 pour évaluer si la perte des cellules souches du bulge a une valeur diagnostique dans l’alopécie cicatricielle et nous avons démontré que cette perte s’observait de manière identique dans le lichen plan pilaire, l’alopécie frontale fibrosante comme dans le lupus érythémateux et n’avait donc aucune valeur diagnostique. Nous avons tenté d’intégrer les nouveaux concepts et nos données dans les classifications traditionnelles des alopécies et nous avons élaboré un nouvel algorithme diagnostique. L’association des immunomarquages avec la technique HoVert ouvre de nouvelles perspectives dans le diagnostic histopathologique des alopécies primaires du cuir chevelu.
Doctorat en Sciences médicales (Médecine)
info:eu-repo/semantics/nonPublished

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Holtzhausen, Wendy. "Diuretic factors controlling beetle malphighian tubules fluid secretion and immunohistochemistry /." Pretoria : [s.n.], 2006. http://upetd.up.ac.za/thesis/available/etd-07182007-164904.

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Fung, Hau-yin Kevin. "Detection of SARS-CoV in lung tissues by immunohistochemistry and in-situ hybridization /." View the Table of Contents & Abstract, 2005. http://sunzi.lib.hku.hk/hkuto/record/B32020624.

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Книги з теми "Immunohistochemistry"

Kalyuzhny, Alexander E. Immunohistochemistry. Cham: Springer International Publishing, 2016. http://dx.doi.org/10.1007/978-3-319-30893-7.

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Simon, Renshaw, ed. Immunohistochemistry. Bloxham: Scion, 2007.

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Chu, Peiguo. Modern immunohistochemistry. Cambridge: Cambridge University Press, 2009.

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C, Cuello A., ed. Immunohistochemistry II. Chichester: Wiley, 1993.

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Del Valle, Luis, ed. Immunohistochemistry and Immunocytochemistry. New York, NY: Springer US, 2022. http://dx.doi.org/10.1007/978-1-0716-1948-3.

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Kalyuzhny, Alexander E., ed. Signal Transduction Immunohistochemistry. New York, NY: Springer New York, 2017. http://dx.doi.org/10.1007/978-1-4939-6759-9.

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Kalyuzhny, Alexander E., ed. Signal Transduction Immunohistochemistry. Totowa, NJ: Humana Press, 2011. http://dx.doi.org/10.1007/978-1-61779-024-9.

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Kalyuzhny, Alexander E., ed. Signal Transduction Immunohistochemistry. New York, NY: Springer US, 2023. http://dx.doi.org/10.1007/978-1-0716-2811-9.

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A, DeLellis Ronald, ed. Advances in immunohistochemistry. New York: Raven Press, 1988.

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Lin, Fan, Jeffrey W. Prichard, Haiyan Liu, and Myra L. Wilkerson, eds. Handbook of Practical Immunohistochemistry. Cham: Springer International Publishing, 2022. http://dx.doi.org/10.1007/978-3-030-83328-2.

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Частини книг з теми "Immunohistochemistry"

Ramos-Vara, José A., and Luke B. Borst. "Immunohistochemistry." In Tumors in Domestic Animals, 44–87. Hoboken, NJ, USA: John Wiley & Sons, Inc., 2016. http://dx.doi.org/10.1002/9781119181200.ch3.

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Clausen, P. P., M. Møller, B. van Deurs, and O. W. Petersen. "Immunohistochemistry." In Theory and Strategy in Histochemistry, 367–86. Berlin, Heidelberg: Springer Berlin Heidelberg, 1991. http://dx.doi.org/10.1007/978-3-642-73742-8_26.

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Chiaravalli, Anna Maria. "Immunohistochemistry." In Encyclopedia of Pathology, 1–5. Cham: Springer International Publishing, 2020. http://dx.doi.org/10.1007/978-3-319-28845-1_5085-1.

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Kigawa, Junzo, Tsunehisa Kaku, Toru Sugiyama, and Steven G. Silverberg. "Immunohistochemistry." In Atlas of Clear Cell Carcinoma of the Ovary, 35–38. Tokyo: Springer Japan, 2015. http://dx.doi.org/10.1007/978-4-431-55438-7_6.

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Azumi, Norio, and Bernard Czernobilsky. "Immunohistochemistry." In Blaustein’s Pathology of the Female Genital Tract, 1131–59. New York, NY: Springer New York, 1994. http://dx.doi.org/10.1007/978-1-4757-3889-6_26.

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Taylor, Clive R., and Richard J. Cote. "Immunohistochemistry." In Encyclopedia of Cancer, 1–6. Berlin, Heidelberg: Springer Berlin Heidelberg, 2015. http://dx.doi.org/10.1007/978-3-642-27841-9_2995-2.

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Baron, Jeffrey, Jeffrey M. Voigt, Thomas T. Kawabata, and Jan A. Redick. "Immunohistochemistry." In Regulation of Hepatic Metabolism, 87–118. Boston, MA: Springer US, 1986. http://dx.doi.org/10.1007/978-1-4684-5041-5_4.

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Davis, Barbara J. "Immunohistochemistry." In Encyclopedia of Systems Biology, 1002. New York, NY: Springer New York, 2013. http://dx.doi.org/10.1007/978-1-4419-9863-7_1594.

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Carbone, Giancarlo. "Immunohistochemistry." In Antibody Usage in the Lab, 84–124. Berlin, Heidelberg: Springer Berlin Heidelberg, 1999. http://dx.doi.org/10.1007/978-3-662-03942-7_6.

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Chiaravalli, Anna Maria. "Immunohistochemistry." In Endocrine Pathology, 422–26. Cham: Springer International Publishing, 2022. http://dx.doi.org/10.1007/978-3-030-62345-6_5085.

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Тези доповідей конференцій з теми "Immunohistochemistry"

Hang Chang, Rosa Anna DeFilippis, Thea D. Tlsty, and Bahrain Parvin. "Scoring histological sections through immunohistochemistry." In 2008 5th IEEE International Symposium on Biomedical Imaging (ISBI 2008). IEEE, 2008. http://dx.doi.org/10.1109/isbi.2008.4541003.

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Solic, Ivana, and Katarina Vukojevic. "In Search of Immunohistochemistry Quantification." In 2021 6th International Conference on Smart and Sustainable Technologies (SpliTech). IEEE, 2021. http://dx.doi.org/10.23919/splitech52315.2021.9566462.

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Razzaq, Hasanain H., Rozaida Ghazali, and Loay E. George. "Immunohistochemistry BC image analysis: A review." In PROCEEDINGS OF THE 1ST INTERNATIONAL CONFERENCE ON FRONTIER OF DIGITAL TECHNOLOGY TOWARDS A SUSTAINABLE SOCIETY. AIP Publishing, 2023. http://dx.doi.org/10.1063/5.0113534.

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Wemmert, Cedric, Juliane M. Kruger, Germain Forestier, Ludovic Sternberger, Friedrich Feuerhake, and Pierre Gancarski. "Stain unmixing in brightfield multiplexed immunohistochemistry." In 2013 20th IEEE International Conference on Image Processing (ICIP). IEEE, 2013. http://dx.doi.org/10.1109/icip.2013.6738232.

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Gilbert, B. "Immunohistochemistry for the MEPHISTO X-PEEM." In Sixth international conference on x-ray microscopy (XRM99). AIP, 2000. http://dx.doi.org/10.1063/1.1291142.

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Bartlett, JMS, M. Gustavson, D. Stocken, D. Rimm, J. Christiansen, CJH van de Velde, A. Hasenburg, et al. "Abstract P4-08-02: A Comparison between AQUA Quantitative Fluorescent Immunohistochemistry and Conventional Immunohistochemistry for Hormone Receptors." In Abstracts: Thirty-Third Annual CTRC‐AACR San Antonio Breast Cancer Symposium‐‐ Dec 8‐12, 2010; San Antonio, TX. American Association for Cancer Research, 2010. http://dx.doi.org/10.1158/0008-5472.sabcs10-p4-08-02.

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Chen, Ting, and Chukka Srinivas. "Structure preserving color deconvolution for immunohistochemistry images." In SPIE Medical Imaging, edited by Metin N. Gurcan and Anant Madabhushi. SPIE, 2015. http://dx.doi.org/10.1117/12.2079893.

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Novrial, Dody, Kamal Agung Wijayana, and Hanif Kun Cahyani. "Immunohistochemistry of KRAS Protein in Colorectal Cancer." In 1’s t Jenderal Soedirman International Medical Conference (JIMC) in conjunction with the Annual Scientific Meeting (Temilnas) Consortium of Biomedical Science Indonesia (KIBI ). SCITEPRESS - Science and Technology Publications, 2020. http://dx.doi.org/10.5220/0010487500470051.

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Chang, Young Hwan, Takahiro Tsujikawa, Adam Margolin, Lisa M. Coussens, and Joe W. Gray. "Multiplexed immunohistochemistry image analysis using sparse coding." In 2017 39th Annual International Conference of the IEEE Engineering in Medicine and Biology Society (EMBC). IEEE, 2017. http://dx.doi.org/10.1109/embc.2017.8037744.

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Shafique, Abubakr, Morteza Babaie, Ricardo Gonzalez, and H. R. Tizhoosh. "Immunohistochemistry Biomarkers-Guided Image Search for Histopathology." In 2023 45th Annual International Conference of the IEEE Engineering in Medicine & Biology Society (EMBC). IEEE, 2023. http://dx.doi.org/10.1109/embc40787.2023.10340099.

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Звіти організацій з теми "Immunohistochemistry"

Pleva, Christina M., Tracey A. Hamilton, John P. Petrali, and Robert K. Kan. Determining Optimal Microwave Antigen Retrieval Conditions for Microtubule-Associated Protein 2 Immunohistochemistry in the Guinea Pig Brain. Fort Belvoir, VA: Defense Technical Information Center, December 2002. http://dx.doi.org/10.21236/ada417833.

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Burke, Allen P. Detection and Clinicopathologic Correlation of Human Immunodeficiency Virus (HIV-1) Nucleic Acids and Antigens in Reticuloendothelial and Central Nervous System Tissues, by Immunohistochemistry, in situ Hybridization, and Polymerase Chain Reaction. Fort Belvoir, VA: Defense Technical Information Center, September 1992. http://dx.doi.org/10.21236/ada259307.

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Funkenstein, Bruria, and Cunming Duan. GH-IGF Axis in Sparus aurata: Possible Applications to Genetic Selection. United States Department of Agriculture, November 2000. http://dx.doi.org/10.32747/2000.7580665.bard.

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Many factors affect growth rate in fish: environmental, nutritional, genetics and endogenous (physiological) factors. Endogenous control of growth is very complex and many hormone systems are involved. Nevertheless, it is well accepted that growth hormone (GH) plays a major role in stimulating somatic growth. Although it is now clear that most, if not all, components of the GH-IGF axis exist in fish, we are still far from understanding how fish grow. In our project we used as the experimental system a marine fish, the gilthead sea bream (Sparus aurata), which inhabits lagoons along the Mediterranean and Atlantic coasts of Europe, and represents one of the most important fish species used in the mariculture industry in the Mediterranean region, including Israel. Production of Sparus is rapidly growing, however, in order for this production to stay competitive, the farming of this fish species has to intensify and become more efficient. One drawback, still, in Sparus extensive culture is that it grows relatively slow. In addition, it is now clear that growth and reproduction are physiological interrelated processes that affect each other. In particular sexual maturation (puberty) is known to be closely related to growth rate in fish as it is in mammals, indicating interactions between the somatotropic and gonadotropic axes. The goal of our project was to try to identify the rate-limiting components(s) in Sparus aurata GH-IGF system which might explain its slow growth by studying the ontogeny of growth-related genes: GH, GH receptor, IGF-I, IGF-II, IGF receptor, IGF-binding proteins (IGFBPs) and Pit-1 during early stages of development of Sparus aurata larvae from slow and fast growing lines. Our project was a continuation of a previous BARD project and could be divided into five major parts: i) obtaining additional tools to those obtained in the previous project that are necessary to carry out the developmental study; ii) the developmental expression of growth-related genes and their cellular localization; iii) tissue-specific expression and effect of GH on expression of growth-related genes; iv) possible relationship between GH gene structure, growth rate and genetic selection; v) the possible role of the IGF system in gonadal development. The major findings of our research can be summarized as follows: 1) The cDNAs (complete or partial) coding for Sparus IGFBP-2, GH receptor and Pit-1 were cloned. Sequence comparison reveals that the primary structure of IGFBP-2 protein is 43-49% identical to that of zebrafish and other vertebrates. Intensive efforts resulted in cloning a fragment of 138 nucleotides, coding for 46 amino acids in the proximal end of the intracellular domain of GH receptor. This is the first fish GH receptor cDNA that had been cloned to date. The cloned fragment will enable us to complete the GH - receptor cloning. 2) IGF-I, IGF-II, IGFBP-2, and IGF receptor transcripts were detected by RT-PCR method throughout development in unfertilized eggs, embryos, and larvae suggesting that these mRNAs are products of both the maternal and the embryonic genomes. Preliminary RT-PCR analysis suggest that GH receptor transcript is present in post-hatching larvae already on day 1. 3) IGF-1R transcripts were detected in all tissues tested by RT-PCR with highest levels in gill cartilage, skin, kidney, heart, pyloric caeca, and brain. Northern blot analysis detected IGF receptor only in gonads, brain and gill cartilage but not in muscle; GH increased slightly brain and gill cartilage IGF-1R mRNA levels. 4) IGFBP-2 transcript were detected only in liver and gonads, when analyzed by Northern blots; RT-PCR analysis revealed expression in all tissues studied, with the highest levels found in liver, skin, gonad and pyloric caeca. 5) Expression of IGF-I, IGF-II, IGF-1R and IGFBP-2 was analyzed during gonadal development. High levels of IGF-I and IGFBP-2 expression were found in bisexual young gonads, which decreased during gonadal development. Regardless of maturational stage, IGF-II levels were higher than those of IGF-L 6) The GH gene was cloned and its structure was characterized. It contains minisatellites of tandem repeats in the first and third introns that result in high level of genetic polymorphism. 7) Analysis of the presence of IGF-I and two types of IGF receptor by immunohistochemistry revealed tissue- and stage-specific expression during larval development. Immunohistochemistry also showed that IGF-I and its receptors are present in both testicular and ovarian cells. Although at this stage we are not able to pinpoint which is the rate-limiting step causing the slow growth of Sparus aurata, our project (together with the previous BARD) yielded a great number of experimental tools both DNA probes and antibodies that will enable further studies on the factors regulating growth in Sparus aurata. Our expression studies and cellular localization shed new light on the tissue and developmental expression of growth-related genes in fish.

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Fields, Michael J., Mordechai Shemesh, and Anna-Riitta Fuchs. Significance of Oxytocin and Oxytocin Receptors in Bovine Pregnancy. United States Department of Agriculture, August 1994. http://dx.doi.org/10.32747/1994.7568790.bard.

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Oxytocin has multiple actions in bovine reproductive tract and it was our purpose to determine the nature of these actions and their significance for the physiology of bovine reproduction. The bovine oxytocin receptors (OTR) gene was cloned and its expression studied during the cycle and pregnancy. OTR mRNA changed in parallel with OTR with control occurring mainly at the transcriptional level. However, the endocrine regulation of OTR were found in endometrium and cervical mucosa at estrus and at parturition. In both tissues OTR were suppressed in the luteal phase and early pregnancy. Whereas cervical OTR remained suppressed throughout pregnancy, endometrial OTR began to increase soon after implantation and reached higher concentrations in midpregnancy than at estrus. OTR in caruncles did not increase until third trimester, and OTR in cervical mucosa, cotyledons and fetal membranes increased only at term. Myometrial OTR showed less variation and OTR were present throughout the cycle and pregnancy but increased significantly during mid- and late pregnancy. OTR were localized in endometrial epithelial cells and lumina epithelial cells of cervical mucosa as determined by immunohistochemistry. Endometrial OTR were functional throughout pregnancy and mediated PGF release from day 50 onwards in a receptor density related manner. OTR in cervical mucosa mediated PGE release both in vivo and in vitro, as shown in cyclic cows. The ontogeny of uterine OTR was studied from third trimester fetal stage until puberty. OTR were present in endometrium and cervical mucosa in high concentrations throughout this period; myometrial OTR began to increase somewhat later but also reached adult values by 6-mo of age. In the prepuberal heifers OT injections failed to initiate PGF2a, release. The influence of steroids on the effect of OT was examined. Ovariectomy and E2 were without effect, but P4 with or without E2 induced a massive PGF2a release in response to OT in spite of reduced OTR. Bovine cyclooxygenases (COX-1 and COX-2) were cloned and their expression studied in the endometrium of prepuberal heifers and pregnant cows. Untreated and E2 treated prepuberal heifers did not express COX-2 but P4 treated heifers did express the mRNA for COX-2, albeit weakly. During the second half of pregnancy COX-2 mRNA was strongly expressed in cotyledons and somewhat less in caruncles, whereas endometrium, myometrium and cervical mucosa showed only weak, if any, COX-2 mRNA under basal conditions. However, 2 h after OT injection significant increases in COX-2 mRNA were found in endometrial RNA. Thus OT is capable of inducing the expression of the inducible COX-2 gene, and hence the conversion of arachidonic acid to prostanoids. The results indicate that the functions of OT are numerous and probably essential for successful pregnancy and parturition.

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