EnVision+, a New Dextran Polymer-based Signal Enhancement Technique for In Situ Hybridization (ISH)

Wiedorn, Klaus Hermann; Goldmann, Torsten; Henne, C; Kühl, Heike; Vollmer, Ekkehard

doi:10.1177/002215540104900901

Cited by 15 publications

(6 citation statements)

References 6 publications

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“…This method uses a compact polymer to which 4 to 70 molecules of enzyme (peroxidase or alkaline phosphatase) plus 1 to 10 molecules of the secondary antibody are attached (Table 5). 149,411 Its advantages are (1) simplicity compared with the 3-step methods, (2) equal or higher sensitivity than ABC or LSAB methods, and (3) lack of background due to endogenous biotin or avidin. 269,318,335,370,398 However, this method is usually more expensive than ABC or LSAB methods (Fig.…”

Section: Avidin-biotin Methodsmentioning

confidence: 99%

When Tissue Antigens and Antibodies Get Along

2013

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Once focused mainly on the characterization of neoplasms, immunohistochemistry (IHC) today is used in the investigation of a broad range of disease processes with applications in diagnosis, prognostication, therapeutic decisions to tailor treatment to an individual patient, and investigations into the pathogenesis of disease. This review addresses the technical aspects of immunohistochemistry (and, to a lesser extent, immunocytochemistry) with attention to the antigen-antibody reaction, optimal fixation techniques, tissue processing considerations, antigen retrieval methods, detection systems, selection and use of an autostainer, standardization and validation of IHC tests, preparation of proper tissue and reagent controls, tissue microarrays and other high-throughput systems, quality assurance/quality control measures, interpretation of the IHC reaction, and reporting of results. It is now more important than ever, with these sophisticated applications, to standardize the entire IHC process from tissue collection through interpretation and reporting to minimize variability among laboratories and to facilitate quantification and interlaboratory comparison of IHC results.

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Section: Avidin-biotin Methodsmentioning

confidence: 99%

When Tissue Antigens and Antibodies Get Along

2013

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“…To reduce background staining from endogenous biotin, the EnVision technique was used. 38 The stained sections were then reviewed by three pathologists, and a score was given for percentage of cell staining, strength of staining (1+ to 3+, 3+ being the strongest), and pattern of staining (membranous, cytoplasmic, or nuclear).…”

Section: Methodsmentioning

confidence: 99%

The Immunohistochemical Profile of Malignant Mesotheliomas of the Tunica Vaginalis

Winstanley¹,

Landon²,

Berney³

et al. 2006

The American Journal of Surgical Pathology

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Malignant mesotheliomas of the testis arise from the tunica vaginalis, formed from the evagination of the abdominal peritoneum into the scrotum. The immunohistochemical profile of the tunica vaginalis and associated neoplasms is often extrapolated from thoracic studies. Testicular series are uncommon, usually derived from previous case studies and literature reviews. The immunohistochemical findings in 20 cases originally diagnosed as malignant mesotheliomas are presented. Archival testicular malignant mesothelioma specimens from 1959 to 2004 were collected from hospitals throughout the United Kingdom and from the authors' own archives. Hematoxylin and eosin-stained sections were reviewed, and selected sections from each case were then examined using an immunohistochemical panel of eight antibodies: calretinin (Zymed, 1:200), epithelial membrane antigen (EMA) (DAKO, 1:50), thrombomodulin (DAKO 1:5), CK7 (DAKO, 1:100), CK5-CK6 (DAKO, 1:10), BerEp4 (DAKO, 1:25), carcinoembryonic antigen (CEA) (DAKO, 1:10), and CK20 (DAKO, 1:100). The EnVision technique was used for all antibodies. Sections were reviewed independently by three pathologists. Electron microscopy was performed on selected cases. In all cases, the morphologic light microscopy criteria for a diagnosis of malignant mesothelioma were present. However, two tumors were later excluded from the study because of diffuse strong positive immunostaining with CK20 and BerEp4 and an ultrastructural appearance of adenocarcinoma. Of the remaining cases, 15 of 18 (83%) were purely epithelioid in type, showing a mixture of papillary, tubular, and solid patterns, and 3 of 18 (17%) showed a mixed sarcomatoid/epithelioid pattern. All cases were positive for calretinin and EMA (100%), 16 of 18 (89%) were positive for thrombomodulin, and 15 of 18 (83%) were positive for CK7. CK5-CK6 positivity was present in 13 of 18 (72%) but varied in strength and distribution; 2 of 18 (11%) were positive for BerEp4. All the cases were negative for CK20 and CEA. Four of the 18 cases were examined by electron microscopy, which revealed long thin microvilli supporting a diagnosis of malignant mesothelioma. This study has shown that the immunocytochemical profile of testicular malignant mesotheliomas is similar to those arising in the pleura, with diffuse positivity for calretinin, EMA, thrombomodulin, and CK7, and negative for CK20 and CEA. Focal weak positivity may be encountered with BerEp4. However, histopathologists should be aware of the variability in CK5-6 staining in testicular specimens when compared with pleural mesotheliomas.

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“…The first step is the unlabeled primary antibody; the second step consists of a polymer containing numerous secondary antibodies as well as numerous molecules of enzyme. 59,75 These methods are faster than most current avidin-biotinbased methods, do not produce the background generated by endogenous biotin, and have comparable or sometimes superior sensitivity to avidin-biotin methods. They are also more expensive.…”

Section: Practical Approach To Standardization Of a New Antibodymentioning

confidence: 99%

Suggested Guidelines for Immunohistochemical Techniques in Veterinary Diagnostic Laboratories

et al. 2008

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Abstract. This document is the consensus of the American Association of Veterinary Laboratory Diagnosticians (AAVLD) Subcommittee on Standardization of Immunohistochemistry on a set of guidelines for immunohistochemistry (IHC) testing in veterinary laboratories. Immunohistochemistry is a powerful ancillary methodology frequently used in many veterinary laboratories for both diagnostic and research purposes. However, neither standardization nor validation of IHC tests has been completely achieved in veterinary medicine. This document addresses both issues. Topics covered include antibody selection, fixation, antigen retrieval, antibody incubation, antibody dilutions, tissue and reagent controls, buffers, and detection systems. The validation of an IHC test is addressed for both infectious diseases and neoplastic processes. In addition, storage and handling of IHC reagents, interpretation, quality control and assurance, and troubleshooting are also discussed. Proper standardization and validation of IHC will improve the quality of diagnostics in veterinary laboratories.

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EnVision+, a New Dextran Polymer-based Signal Enhancement Technique for In Situ Hybridization (ISH)

Cited by 15 publications

References 6 publications

When Tissue Antigens and Antibodies Get Along

When Tissue Antigens and Antibodies Get Along

The Immunohistochemical Profile of Malignant Mesotheliomas of the Tunica Vaginalis

Suggested Guidelines for Immunohistochemical Techniques in Veterinary Diagnostic Laboratories

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