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Ish iview blue detection kit

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The ISH iVIEW Blue detection kit is a laboratory equipment product designed for in situ hybridization (ISH) analysis. The kit provides the necessary reagents and components to enable the visualization of target nucleic acid sequences in tissue samples.

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Lab products found in correlation

Inform eber probe, by Roche (4 mentions) Nuclear fast red, by Roche (2 mentions) Benchmark ultra, by Roche (2 mentions) Benchmark xt, by Roche (1 mentions) Red counterstain 2, by Roche (1 mentions) Mum1p, by Agilent Technologies (1 mentions) Tma master 01, by 3DHISTECH (1 mentions) Ber h2, by Agilent Technologies (1 mentions) Autostainer, by Roche (1 mentions) Ventana benchmark, by Roche (1 mentions) Bond max autostainer, by Leica (1 mentions) Benchmark xt autostainer, by Roche (1 mentions)

Close spelling variants of this product

Ventana ISH iVIEW Blue Detection Kit ISH iVIEW Blue Plus Detection Kit Ventana ISH iView Blue Plus detection kit ISH iView kit

8 protocols using ish iview blue detection kit

1

EBV Status Determination by ISH

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EBV status was determined by Epstein-Barr virus-encoded Early RNA (EBER) in situ hybridization (ISH) on the samples, brushes or TMA. Ventana BenchMark automated staining instruments (Ventana Medical systems, Tuscon, AZ, USA) were used for ISH of the samples or TMA using an EBV-specific probe (INFORM EBER PROBE; Ventana Medical systems) and ISH iVIEW Blue detection kit (Ventana Medical systems, Inc.) for staining using the manufacturer’s instructions in Innsbruck, Utrecht, Hong Kong, Stanford and London. Shenzhen used an EBER Probe (Zhongshan Jinquaiao Biotechnology Co.; Beijing, China) and an autostainer (Ventana Medical Systems, Inc.) to perform ISH. Singapore used a BONDTM Ready-to-use ISH EBER probe and a Leica Bond-Max autostainer (all Leica Biosystems, Wetzlar, Germany) for this purpose. In situ hybridization of xenografts and cell pellets was done using an EBV-specific probe (INFORM EBER PROBE; Ventana Medical systems) and ISH iVIEW Blue detection kit (Ventana Medical systems, Inc.) using the manufacturer’s instructions.
Lechner M., Schartinger V.H., Steele C.D., Nei W.L., Ooft M.L., Schreiber L.M., Pipinikas C.P., Chung G.T., Chan Y.Y., Wu F., To K.F., Tsang C.M., Pearce W., Morelli D., Philpott M., Masterson L., Nibhani R., Wells G., Bell C.G., Koller J., Delecluse S., Yip Y.L., Liu J., Forde C.T., Forster M.D., Jay A., Dudás J., Krapp A., Wan S., Uprimny C., Sprung S., Haybaeck J., Fenton T.R., Chester K., Thirlwell C., Royle G., Marafioti T., Gupta R., Indrasari S.R., Herdini C., Slim M.A., Indrawati I., Sutton L., Fles R., Tan B., Yeong J., Jain A., Han S., Wang H., Loke K.S., He W., Xu R., Jin H., Cheng Z., Howard D., Hwang P.H., Le Q.T., Tay J.K., West R.B., Tsao S.W., Meyer T., Riechelmann H., Oppermann U., Delecluse H.J., Willems S.M., Chua M.L., Busson P., Lo K.W., Wollmann G., Pillay N., Vanhaesebroeck B, & Lund V.J. (2021). Somatostatin receptor 2 expression in nasopharyngeal cancer is induced by Epstein Barr virus infection: impact on prognosis, imaging and therapy. Nature Communications, 12, 117.
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2

EBER-ISH Staining Procedure for Detection

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Biopsy specimens were fixed in formalin, dehydrated, embedded in paraffin, and sectioned with routine methods. The INFORM EBER probe (Ventana Medical Systems, Tucson, AZ, USA) was used for EBER‐ISH. Slides were stained on an automated stainer (Benchmark XT; Ventana Medical Systems), and for visualization the ISH iView Blue Detection Kit (Ventana Medical Systems) with alkaline phosphatase and NBT/BCIP substrate was used, with Nuclear Fast Red (Ventana Medical Systems) for contrast. Specimens in which nuclear expression of EBER was observed in 20% or more of the malignant cells were considered EBER‐positive.15
Okamoto A., Yanada M., Miura H., Inaguma Y., Tokuda M., Morishima S., Kanie T., Yamamoto Y., Mizuta S., Akatsuka Y., Yoshikawa T., Mizoguchi Y., Nakamura S., Okamoto M, & Emi N. (2015). Prognostic significance of Epstein–Barr virus DNA detection in pretreatment serum in diffuse large B‐cell lymphoma. Cancer Science, 106(11), 1576-1581.
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3

Detection of EBV by ISH Staining

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According to the manufacturer’s instruction, EBV was detected by in-situ-hybridization (ISH) using the INFORM EBER probe (Ventana Medical Systems, Inc., Tucson, AZ, USA). The slides were stained using an automated stainer (Benchmark Ultra, Ventana Medical Systems, Inc., Tucson, AZ, USA). The visualization was by ISH iView Blue Detection kit (Ventana Medical Systems, Inc., Tucson, AZ, USA) with alkaline phosphatase 5-bromo-4-chloro-3-indolyl phosphate/nitro blue tetrazolium (BCIP NBT/) substrate. Nuclear Fast red (Ventana Medical Systems, Inc., Tucson, AZ, USA) was used as a contrast. Appropriate positive and negative controls were set up parallel to the section analyzed.
Uzoma I.C., Taiwo I.A., Granai M., Di Stefano G., Sorrentino E., Mannucci S., Durosinmi M.A., Lazzi S., Leoncini L, & Akinloye O. (2021). Distinct pattern of lymphoid neoplasms characterizations according to the WHO classification (2016) and prevalence of associated Epstein–Barr virus infection in Nigeria population. Infectious Agents and Cancer, 16, 36.
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4

EBER-ISH Analysis of FFPE Tissue Samples

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Retrieved FFPE tissue, was sectioned and slides prepared. All histological slides were reviewed and classified according to the World Health Organization (WHO) Classification of Head and Neck Tumours (2017 edition)29 . EBER-ISH for EBER1 and EBER2 were performed, for cases where this information was not already available, on four µm thick sections using an INFORM EBER Probe with ISH iVIEW Blue Detection Kit on BenchMark Ultra (all Ventana Medical Systems, Tucson, AZ). Appropriate positive specimen controls were used as well as Negative Control Probe (Ventana Medical Systems), the latter for assessment of non-specific background staining. The pathologist (F.C.A.) was unaware of the outcome of EBV-DNA analyses, while performing the classification. The methodology used for EBER analysis was the same as used for the historic cases. It has previously been demonstrated that EBERs are stable in formalin-fixed paraffin-embedded tissues30 (link), and no difficulties were experienced with the present analysis.
Nilsson J.S., Forslund O., Andersson F.C., Lindstedt M, & Greiff L. (2019). Intralesional EBV-DNA load as marker of prognosis for nasopharyngeal cancer. Scientific Reports, 9, 15432.
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5

EBER In Situ Hybridization for EBV Detection

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EBV-encoded RNA (EBER) in situ hybridization was performed with INFORM EBER Probes (a probe used for the detection of early RNA transcript of EBV infection, Catalog number 800-2842, Ventana, Tucson, AZ, USA) and Ventana Medical Systems' ISH iVIEWBlue Detection Kit (Catalog number 800-092, Ventana, Tucson, AZ, USA) on 4 μm-thick formalin-fixed, paraffin-embedded sections. The reaction was detected using Ventana Medical Systems' Red CounterstainII (Catalog number 780-2218, Ventana, Tucson, AZ, USA). A case of EBV+ nasopharyngeal carcinoma (NPC) was used as a positive control.
Wang L., Dong H., Ni S., Huang D., Tan C., Chang B, & Sheng W. (2016). Programmed death-ligand 1 is upregulated in intrahepatic lymphoepithelioma-like cholangiocarcinoma. Oncotarget, 7(43), 69749-69759.
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6

Immunohistochemical Profiling of PTLD Samples

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Primary diagnostic formalin fixed paraffin-embedded tissue samples containing representative PTLD were available from 52 patients; samples of these were included in a tissue microarray (TMA). Briefly, three 1-mm diameter tissue cores were identified in tumor areas, punched out and re-embedded in recipient blocks using a TMA master-01 (3DHISTECH Ltd., Budapest, Hungary).
Immunohistochemical stains were performed on 4 μm formalin fixed paraffin-embedded tissue sections according to standard antibody-specific protocols, optimized in house for use with the Ventana Benchmark automated staining system (Ventana Medical Systems, Tucson, AZ). For review of the classification of the tumors according to WHO 2008 criteria, a panel of markers was applied consisting of CD3 (polyclonal, Dako, Glostrup, Denmark), CD5 (SP19, Ventana Medical Systems), CD4 (SP35, Ventana), CD8 (SP57, Ventana), CD10 (SP67, Ventana), CD20 (L26, Ventana), CD79a (MRQ-48, Ventana), CD30 (Ber-H2, Dako) and MUM1 (MUM1P, Dako). The EBV antigens were visualized using antibodies to LMP (LMP clones CS.1-4, Dako) and EBNA2 (PE2; Abcam, Cambridge, UK). EBV-RNA (EBER) expression was identified by in-situ hybridization (ISH iView Blue Detection kit; Ventana). In each case, EBV status in the PTLD cell population was evaluated by the authors (S.H.D. and M.V.) and recorded as either positive or negative.
Vase M.Ø., Maksten E.F., Strandhave C., Søndergaard E., Bendix K., Hamilton-Dutoit S., Andersen C., Møller M.B., Sørensen S.S., Kampmann J., Eiskjær H., Iversen M., Weinreich I.D., Møller B., Jespersen B, & d'Amore F. (2015). HLA Associations and Risk of Posttransplant Lymphoproliferative Disorder in a Danish Population-Based Cohort. Transplantation Direct, 1(7), e25.
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7

Evaluating Microsatellite Instability in FFPE Tissues

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MSI status was determined by IHC for mismatch repair (MMR) proteins (MLH-1, MSH-2, PMS-2, and MSH-6) in formalin-fixed paraffin-embedded (FFPE) tissues. MSI-high was defined as negative staining for at least one of the MMR proteins. EBV ISH was performed in FFPE tissues using the Bench Mark XT autostainer (Ventana Medical Systems) and ISH iVIEW Blue Detection Kit (Ventana Medical Systems), according to the manufacturer’s instructions.
Kim J.H., Ryu M.H., Park Y.S., Ma J., Lee S.Y., Kim D, & Kang Y.K. (2022). Predictive biomarkers for the efficacy of nivolumab as ≥ 3rd-line therapy in patients with advanced gastric cancer: a subset analysis of ATTRACTION-2 phase III trial. BMC Cancer, 22, 378.
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8

EBER in situ Hybridization Assay

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The EBER in situ hybridization assay was performed on TMA block sections. Epstein–Barr virus (EBV) was identified using EBER probes (ISH iVIEW Blue detection kit; Ventana Medical Systems Inc., Oro Valley, AZ, USA).
Ehara T., Uehara T., Nakajima T., Kinugawa Y., Kobayashi S., Iwaya M., Ota H, & Soejima Y. (2021). LGR5 expression is associated with prognosis in poorly differentiated gastric adenocarcinoma. BMC Cancer, 21, 228.
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