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Benchmark automated immunostainer

Manufactured by Roche
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Sourced in United Kingdom, United States

The Benchmark automated immunostainer is a laboratory equipment product designed for the automated processing of immunohistochemistry (IHC) and in situ hybridization (ISH) assays. It provides consistent and reliable results by automating the staining process, reducing manual handling and potential errors.

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Lab products found in correlation

Pd l1 clone 22c3, by Agilent Technologies (1 mentions) Gata3, by Biocare Medical (1 mentions) Cyclin e, by Santa Cruz Biotechnology (1 mentions) Mrq 50, by Cell Marque (1 mentions)

5 protocols using benchmark automated immunostainer

1

Quantifying Tumor Immune Profiles

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Four-micrometer-thick tumor sections were stained for PD-L1 (clone 22C3, DAKO, Agilent, CA, USA), CD4 (clone BP6028; Biolynx, Hangzhou, China), CD8 (clone: EP334, Abcam, Cambrige, UK) using BenchMark automated immunostainer (Ventana, AZ, USA). Staining was evaluated in a blinded fashion by the pathologists. Scoring was assessed based on the proportion of positive cells among nucleated cells in the ROI. The positive cells were defined as tumor cells displaying membranous staining of PD-L1. The PD-L1 expression at MPC was quantified as the proportion of PD-L1-positive tumor cells in total tumor cells within the MPC area. The tumor proportion score (TPS) is a PD-L1 measurement that has been applied in clinical trials and in clinic of lung cancer (20 (link)). The percentage of CD4+ and CD8+ T cell in the peritumor region was assessed based on the proportion of positive T cells among nucleated cells in the peritumoral area, which was described in previous studies (21 (link), 22 (link)).
Zhang S., Xu Y., Zhao P., Bao H., Wang X., Liu R., Xu R., Xiang J., Jiang H., Yan J., Wu X., Shao Y., Liang J., Wu Q., Zhang Z., Lu S, & Ma S. (2021). Integrated Analysis of Genomic and Immunological Features in Lung Adenocarcinoma With Micropapillary Component. Frontiers in Oncology, 11, 652193.
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2

Immunohistochemical Markers in DLBCL and CHL

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Representative tissue sections that were 4 μm in thickness were used for IHC and EBV-encoded RNA (EBER) in situ hybridization (ISH). The primary antibodies used for routine pathologic examination are: CD3, CD5, CD10, CD15, CD20, CD23, CD30, CD79a, BCL6, Pax5, and IRF4/MUM-1. Additionally, IHC for GATA3 (Biocare Medical, Concord, CA), p63 (Lab Vision, Fremont, CA) and cyclin E (Santacruz Biotechnology, Santa Cruz, CA) was performed to determine their usefulness in the differential diagnosis. Expression of p63, GATA3 and cyclin E was considered positive if the percentage of positive cells was equal to or higher than 5%. To elucidate whether those three proteins are site specific marker, 88 cases of non-mediastinal DLBCL and 56 non-mediastinal CHL were investigated using tissue microarray blocks. All IHC and EBER ISH assays were conducted using the Bench Mark automated immunostainer (Ventana Medical Systems, Tucson AZ) according to validated protocols.
Kim H.J., Kim H.K., Park G., Min S.K., Cha H.J., Lee H., Choi S.J., Na H.Y., Choe J.Y, & Kim J.E. (2019). Comparative pathologic analysis of mediastinal B-cell lymphomas: selective expression of p63 but no GATA3 optimally differentiates primary mediastinal large B-cell lymphoma from classic Hodgkin lymphoma. Diagnostic Pathology, 14, 133.
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3

NMZL Identification Protocol: Pathological Evaluation

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Cases of NMZL were identified from the files at the National Cancer Institute and the Cancer University Institute of Toulouse Oncopole. Additional cases were contributed by coauthors as members of the International Lymphoma Study Group, reviewed by an expert panel (E. S.J., E.C., S.P., S.H.S.), with a consensus diagnosis of NMZL reached. The study was approved by the Institutional Review Board of the National Cancer Institute.
Hematoxylin and eosin (H&E)-stained sections from each case were evaluated. Histology reports, available flow cytometry results, and immunohistochemistry slides were reviewed. Additional immunohistochemical studies were performed on available material with CD20, CD3, CD79a, CD10, BCL6, MUM1 (IRF4), CD4, CD8, PD1, ICOS, CXCL13, FOXP3, PDL1, CD21, IgD, kappa, and lambda. Immunohistochemistry was performed on a Ventana Benchmark automated immunostainer using UltraView detection. The panel of antibodies, clones, and sources are presented in Table 1.
The cytologic composition of the cases was evaluated and the presence of the following cells types recorded: centrocyte-like cells, monocytoid cells, and plasmacytoid cells/plasma cells. The morphology of these cell types has been described elsewhere.2 (link) Plasmacytoid differentiation required the presence of light-chain restricted plasmacytoid cells/plasma cells by immunohistochemistry.
Egan C., Laurent C., Alejo J.C., Pileri S., Campo E., Swerdlow S.H., Piris M., Chan W.C., Warnke R., Gascoyne R.D., Xi L., Raffeld M., Pittaluga S, & Jaffe E.S. (2020). Expansion of PD1-positive T Cells in Nodal Marginal Zone Lymphoma: A Potential Diagnostic Pitfall. The American journal of surgical pathology, 44(5), 657-664.
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4

Immunohistochemical Analysis of Pax8 and Inhibin

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The primary antibodies used were anti-human Pax8 (MRQ-50, prediluted, Cell Marque) and anti-human inhibin (alpha, R1, prediluted, Cell Marque). Immunohistochemical staining for Pax8, and Inhibin was performed on formalin-fixed, paraffin-embedded, 4-µm tissue sections using a Ventana Benchmark automated immunostainer (Ventana, Tucson, AZ, USA). Tissue sections were deparaffinized, and localization of the antigen–antibody complex was achieved using the OptiView DAB detection kit. Antigen retrieval was performed using heat-induced epitope retrieval for 32 min for Pax8, or 64 min for inhibin, in cell conditioning 1 buffer.
Pax8 immunoreactivity was interpreted using nuclear staining alone, whereas inhibin immunoreactivity was interpreted using cytoplasmic staining [2 (link), 10 (link), 11 (link), 19 (link)]. The percentage of Pax8- or inhibin-immunoreactive tumor cells was scored as a proportion. Representative fields were selected and imaged at 400 × magnification using the same microscope. A 5% immunoreactivity cutoff was used to classify a case as positive.
Chi Z., Xu J., Karamchandani D.M, & Peng L. (2023). Pax8 as a useful adjunct marker to differentiate pancreatic serous cystadenoma from clear cell renal cell carcinoma in both cytologic and surgical specimens. Diagnostic Pathology, 18, 54.
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5

Immunohistochemical Profiling of Melanocytic Lesions

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Immunohistochemical staining for p16, BRAF V600E, PRAME, and ALK‐1 was performed on the Ventana Medical Systems (VMS) Benchmark Automated Immunostainer (VMS) as previously described.16 p16 expression patterns were categorized similar to prior studies.16 PRAME was designated as positive when greater than 75% of melanocytic nuclei stained positively compared to controls.22, 23 In cases without selected available IHC, stains were performed and blindly scored retrospectively for: p16 (5/134, 4% of cases) BRAF V600E (24/88, 27%), and PRAME (32/56, 57%). Details on processes and reagents are included in Methods S1.
McAfee J.L., Scarborough R., Jia X.S., Azzato E.M., Astbury C., Ronen S., Andea A.A., Billings S.D, & Ko J.S. (2022). Combined utility of p16 and BRAF V600E in the evaluation of spitzoid tumors: Superiority to PRAME and correlation with FISH. Journal of Cutaneous Pathology, 50(2), 155-168.
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