The EnVision++ system: a new immunohistochemical method for diagnostics and research. Critical comparison with the APAAP, ChemMate, CSA, LABC, and SABC techniques

Sabattini, Elena; Bisgaard, Kirsten; Ascani, Stefano; Poggi, Simonetta; Piccioli, Milena; Ceccarelli, Claudio; Pieri, Federica; Orcioni, Giulio Fraternali; Pileri, Stefano

doi:10.1136/jcp.51.7.506

Cited by 396 publications

(286 citation statements)

References 22 publications

(28 reference statements)

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“…No PrP Sc in human systemic amyloid 379 from seven cases, identified by red-green dichroism in cross-polarized light after Congo red staining of adjacent sections, but were not stained by monoclonal antiPrP antibodies 3F4 or KG9 (Figure 1C, E-G) using a highly sensitive immunohistochemical detection system [11,29]. Also, no specific staining was observed elsewhere in these tissues.…”

Section: Is Prp Associated With Human Systemic Amyloid Deposits In Situ?mentioning

confidence: 94%

“…Serial deparaffinized tissue sections (4 µm thick; Superfrost Plus slides; VWR, Poole, UK) were autoclaved at 121 • C for 10 min in distilled water, then pre-treated with 96% v/v formic acid (5 min) and proteinase K (VWR; 10 µg/ml, 5 min) at room temperature. After blocking endogenous tissue biotin (Vector Laboratories, Peterborough, UK), immunohistochemistry with KG9 (1 : 2000 dilution) and 3F4 (1 : 15-750) was performed using the Catalysed Signal Amplification (CSA; DakoCytomation) technique [11,29] according to the manufacturer's recommendations, followed by diaminobenzidine detection and haematoxylin counterstaining. Each batch of immunostaining included positive (CJD brain tissue, including cases with PrP amyloid deposits) and negative (primary antibody substituted with normal human serum) control sections.…”

Section: Ga Tennent Et Almentioning

confidence: 99%

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Disease‐associated prion protein is not detectable in human systemic amyloid deposits

Tennent

Head

Bishop

et al. 2007

The Journal of Pathology

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Cerebral and cardiac amyloid deposits have been reported after scrapie infection in transgenic mice expressing variant prion protein (PrP C ) lacking the glycophosphatidylinositol anchor. The amyloid fibril protein in the systemic amyloid deposits was not characterized, and there is no clinical or pathological association between prion diseases and systemic amyloidosis in humans. Nevertheless, in view of the potential clinical significance of these murine observations, we tested both human amyloidotic tissues and isolated amyloid fibrils for the presence of PrP Sc , the prion protein conformation associated with transmissible spongiform encephalopathy (TSE). We also sequenced the complete prion protein gene, PRNP, in amyloidosis patients. No specific immunohistochemical staining for PrP Sc was obtained in the amyloidotic cardiac and other visceral tissues of patients with different types of systemic amyloidosis. No protease-resistant prion protein, PrP res , was detectable by Western blotting of amyloid fibrils isolated from cardiac and other systemic amyloid deposits. Only the complete normal wild-type PRNP gene sequence was identified, including the usual distribution of codon 129 polymorphisms. These reassuringly negative results do not support the idea that there is any relationship of prions or TSE with human systemic amyloidosis, including cardiac amyloid deposition.

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Section: Is Prp Associated With Human Systemic Amyloid Deposits In Situ?mentioning

confidence: 94%

Section: Ga Tennent Et Almentioning

confidence: 99%

Disease‐associated prion protein is not detectable in human systemic amyloid deposits

Tennent

Head

Bishop

et al. 2007

The Journal of Pathology

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“…Unfortunately, formalin fixation and paraffin embedding of tissue greatly jeopardizes immunohistochemical demonstration of cyclin D1. This hurdle can be overcome by a variety of strategies based on improvement of antigen retrieval, employment of several antibodies, or use of sensitive detection systems such as labeled streptavidin biotin or EnVisionϩ (12)(13)(14)(15)(16)(17). Many immunohistochemistry laboratories, however, continue to obtain weak cyclin D1 signals despite resorting to the aforesaid strategies.…”

Section: Discussionmentioning

confidence: 99%

“…On the other hand, catalyzed signal amplification methods are extremely sensitive and very useful when antigen concentration is below the detection threshold of routine immunohistochemical procedures. Consequently, they are able to yield stronger intensities and significantly improve the results obtained with commercially available anti-cyclin D1 monoclonal antibodies (12,13).…”

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confidence: 99%

Catalyzed Signal Amplification for Cyclin D1 Detection in Mantle Cell Lymphoma

et al. 2003

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Mantle cell lymphoma is characterized by a t(11; 14)(q13;q32) translocation resulting in cyclin D1 protein overexpression. Immunohistochemical detection of the latter, therefore, is a useful marker for the diagnosis of mantle cell lymphoma. Nevertheless, interpretation of results is often hampered by the weak immunoreactivity obtained with routine detection techniques. This problem can be overcome by resorting to highly sensitive catalyzed signal amplification methods based on peroxidasecatalyzed deposition of a biotinylated phenolic compound. The present study compares the results obtained with catalyzed signal amplification, labeled streptavidin biotin, and dextran polymeric conjugate (EnVision؉) techniques in cyclin D1 demonstration in mantle cell lymphoma. The study was performed on formalin-fixed, paraffinembedded archival tissue from 20 mantle cell lymphoma cases. Ten cases of small lymphocytic lymphoma and 10 instances of follicular center cell lymphoma were used as controls. Mantle cell lymphoma, a malignant proliferation of small or medium-sized B lymphocytes that are the neoplastic counterpart of normal lymphoid follicle mantle cells, adopts a variety of growth patterns. This neoplasm is associated with a median survival of 3-5 years and lack of cure in the vast majority of patients. In fact, the 5-year overall survival of mantle cell lymphoma patients (30%) is among the worst for the various lymphoma types (1-7). Distinguishing this more aggressive type of B-cell lymphoma from others (marginal zone, small lymphocytic, and follicular lymphomas) can be very difficult.The most useful mantle cell lymphoma marker is cyclin D1, a cell cycle regulator that stimulates transition from G1 phase to S phase by cooperating with cdk4-6 in Rb phosphorylation and E2F release. Normally, lymphocytes do not express cyclin D1 or produce it in very low, immunohistochemically undetectable levels. In contrast, mantle cell lymphoma lymphocytes show immunohistochemically demonstrable cyclin D1 overexpression as a consequence of a t(11;14)(q13;q32) translocation involving the bcl-1 region of chromosome 11 and the immunoglobulin heavy chain gene of chromosome 14 (8 -10). This translocation is shown by cytogenetics or Southern blotting techniques in 70 -75% of mantle cell lymphoma cases and by fluorescence in situ hybridization (FISH) studies in virtually all mantle cell lymphoma cases (11). As for immuno-

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“…Many double staining combinations can be composed of any mouse monoclonal antibody combined with either a classical rabbit polyclonal antibody or one of the recently introduced commercially available rabbit monoclonal antibodies (Rossi et al 2005). A biotin-free detection system is applied using single species anti-mouse and anti-rabbit polymers attached with either HRP or AP enzymes (Sabattini et al 1998). Because all anti-mouse, anti-rabbit polymers contain a secondary antibody of goat origin, there is no interspecies cross-reaction problem.…”

Section: Mouse-rabbit Combinationmentioning

confidence: 99%

Multiple Immunoenzyme Staining: Methods and Visualizations for the Observation With Spectral Imaging

Loos

2007

J Histochem Cytochem.

190

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Several staining concepts and color combinations exist to perform successful double immunoenzyme staining on human tissue specimens. Most of these concepts are based on differences between both primary antibodies: animal species, mouse Ig isotype or IgG subclasses, conjugates, or concentrations. Traditionally, double immunoenzyme staining has used chromogens selected to provide maximum color contrast when observed with the unaided eye. Unfortunately, visually good color combinations always include at least one diffuse chromogen, because of the paucity of appropriate chromogen colors. This situation is drastically changed with the use of spectral imaging, where multicolor microscopy can be unmixed in individual images based on their spectral characteristics. Spectral unmixing can be performed even up to quadruple immunoenzyme staining. This work contains practical suggestions for immunoenzyme double staining procedures for some frequently encountered primary antibody combinations: rabbit–mouse, goat–mouse, mouse–mouse, and rabbit–rabbit. The suggested protocols are all suitable for a classical red-brown color combination plus blue nuclear counterstain that is composed of peroxidase activity (diaminobenzidine tetrahydrochloride), alkaline phosphatase activity (Liquid Permanent Red), and hematoxylin, respectively. Although the red and brown chromogens do not contrast very well visually, they both show a crisp localization and can be perfectly unmixed by spectral imaging.

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The EnVision++ system: a new immunohistochemical method for diagnostics and research. Critical comparison with the APAAP, ChemMate, CSA, LABC, and SABC techniques

Cited by 396 publications

References 22 publications

Disease‐associated prion protein is not detectable in human systemic amyloid deposits

Disease‐associated prion protein is not detectable in human systemic amyloid deposits

Catalyzed Signal Amplification for Cyclin D1 Detection in Mantle Cell Lymphoma

Multiple Immunoenzyme Staining: Methods and Visualizations for the Observation With Spectral Imaging

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