Discovery xt platform

Manufactured by Roche

Sourced in United States

The Discovery XT Platform is a lab equipment designed for high-throughput screening and analysis. It features automated sample handling and processing capabilities to enable efficient and accurate data generation.

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Lab products found in correlation

Chromomap kit with anti rabbit omnimap, by Roche (5 mentions) Halo software, by Indica Labs (2 mentions) Pannoramic scan 2, by 3DHISTECH (2 mentions) Decloaking chamber, by Biocare Medical (1 mentions) Male athymic nude mice, by Central Lab Animal (1 mentions) D luciferin, by Promega (1 mentions) Ck20 clone ks20, by Agilent Technologies (1 mentions) Dab map kit, by Roche (1 mentions) Anti human cd3, by Agilent Technologies (1 mentions) Anti human cd8 clone c8 144b, by Agilent Technologies (1 mentions) Anti ph3 ser10, by Merck Group (1 mentions) Anti cd44, by BD (1 mentions) Anti olig2, by Merck Group (1 mentions) Anti gfap, by Agilent Technologies (1 mentions) Anti iba1, by Fujifilm (1 mentions)

Close spelling variants of this product

Ventana Discovery XT Autostainer platform (4 citations) Discovery XT biomarker platform (6 citations) Ventana Discovery XT platform (5 citations) Discovery XT (190 citations)

12 protocols using discovery xt platform

Detecting NICD1 in GBM Tissue

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Chromogenic Immunohistochemistry was performed on formalin-fixed paraffin-embedded GBM tissue on a Discovery XT platform (Ventana Medical Systems Inc., Tucson, AZ) using Ventana’s reagents, unless otherwise noted. Six µm-thick sections were deparaffinized in xylene and rehydrated through graded alcohols. Heat induced antigen retrieval was performed in a BioCare Decloaking Chamber in Tris-EDTA buffer for 20 minutes at 120^°C and 17 PSI and incubated in 3% hydrogen peroxide for 4 min. NICD1 antibody (1:100, Abcam, ab8925) was incubated for 3 hours and detected using anti-rabbit horseradish peroxidase multimer (OmniMap). Immune complexes were visualized with 3,3’-diaminobenzidene (DAB) and enhanced with copper sulfate (ChromoMap). Slides were counterstained with hematoxylin, dehydrated and mounted with permanent media.

Bayin N.S., Frenster J.D., Sen R., Si S., Modrek A.S., Galifianakis N., Dolgalev I., Ortenzi V., Illa-Bochaca I., Khahera A., Serrano J., Chiriboga L., Zagzag D., Golfinos J.G., Doyle W., Tsirigos A., Heguy A., Chesler M., Barcellos-Hoff M.H., Snuderl M, & Placantonakis D.G. (2017). Notch signaling regulates metabolic heterogeneity in glioblastoma stem cells. Oncotarget, 8(39), 64932-64953.

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Immunohistochemical Analysis of Phospho-RIPK3

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After collection, embryos and placentas were immersion-fixed in formalin and transferred to phosphate-buffered saline (PBS) before tissue processing. Tissues were then dehydrated in 70% ethanol followed by Flex 95 (Richard-Allan Scientific) and Flex 100 solutions, cleared in xylene, and routinely infiltrated and embedded in paraffin. Embryos and placentas were embedded in separate blocks.
Formalin-fixed, paraffin-embedded tissue sections were stained with rabbit anti-mouse phospho-RIPK3 antibody (5 μg/ml; GEN135–35-9, Genentech) recognizing phosphorylated residues Thr²³¹, as previously described (18 (link)). Immunohistochemistry was performed on the Ventana Discovery XT platform with Cell Conditioning 1 standard antigen retrieval. The reaction was detected with the HQ amplification system (p-RIPK3) using 3,3′ diaminobenzidine as the chromogen and hematoxylin counterstain.

Raju S., Whalen D.M., Mengistu M., Swanson C., Quinn J.G., Taylor S.S., Webster J.D., Newton K, & Shaw A.S. (2018). Kinase domain dimerization drives RIPK3-dependent necroptosis. Science signaling, 11(544), eaar2188.

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Immunohistochemical Tumor Xenograft Analysis

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Xenografted tumors were fixed in 10% neutral buffered formalin, paraffin embedded, and hematoxylin-eosin-saffron (HES) stained. Outgrowths were analyzed by immunohistochemistry (IHC) for the expression of biomarkers. Immunostaining was performed on a Discovery XT Platform (Ventana Medical System, Tucson, AZ, part of Roche Diagnostics) with antigen retrieval using either EDTA buffer, pH 8.0 (CC1, Ventana Medical System) or citrate buffer 10 mM, pH 6.0 (CC2, Ventana Medical System). Primary antibodies were mostly monoclonal rabbit antibodies, and paired slides immunostained with rabbit IgG were used as negative controls. Incubation and color development involved anti-rabbit multimer secondary antibody (horseradish peroxidase complex) with DAB (3,30-diaminobenzidine tetrahydrochloride) as a substrate (ChromoMap Kit with Anti-rabbit OmniMap, Ventana Medical System). The IHC slides were scanned using a Pannoramic SCAN II (3DHISTECH). We then used the HALO software (Indica Labs) to quantify the expression levels of ER, pAkt (S473), and p-S6riboprotein (S235/6).

Jacquemetton J., Kassem L., Poulard C., Dahmani A., De Plater L., Montaudon E., Sourd L., Morisset L., El Botty R., Chateau-Joubert S., Vacher S., Biche I., Treilleux I., Trdan O., Marangoni E, & Le Romancer M. (2021). Analysis of genomic and non-genomic signaling of estrogen receptor in PDX models of breast cancer treated with a combination of the PI3K inhibitor alpelisib (BYL719) and fulvestrant. Breast Cancer Research : BCR, 23, 57.

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Glioblastoma Xenograft Model in Mice

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We used male athymic nude mice (6 wk old; Central Lab. Animal Inc.). The mice were housed in micro-isolator cages under sterile conditions and observed for at least 1 week before study initiation, to ensure proper health. Lighting, temperature, and humidity were centrally controlled. Dissociated GBM TSs (5 × 10⁵ per mouse) were implanted into the right frontal lobe of mice at a depth of 4.5 mm, using guide-screw system [35 (link)]. The mice were randomly allocated based on their body weights without blinding (n = 5 mice per group). If the body weight decreased by more than 15% relative to the maximum weight, mice were euthanized according to the approved protocol. For bioluminescence acquisition and analyses, mice were injected intraperitoneally with 100 μL D-luciferin (30 mg/mL; Promega) under 2.5% isoflurane anesthesia, 15 min before signal acquisition, and then observed using IVIS imaging system and Living Image v4.2 software (Caliper Life Sciences). For immunohistochemistry, sections (5 μm thick) were obtained using a microtome and transferred onto adhesive slides. Antigen retrieval and antibody attachment were performed using the Discovery XT platform (Ventana Medical Systems). Zeb1 was detected using a peroxidase/DAB staining.

Park J., Shim J.K., Lee M., Kim D., Yoon S.J., Moon J.H., Kim E.H., Park J.Y., Chang J.H, & Kang S.G. (2023). Classification of IDH wild-type glioblastoma tumorspheres into low- and high-invasion groups based on their transcriptional program. British Journal of Cancer, 129(7), 1061-1070.

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Immunohistochemical analysis of PKD1 in breast cancer

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Paraffin-embedded breast tumors samples, obtained at the time of initial diagnosis, were retrieved from the archives of the Department of Biopathology at René Huguenin Hospital. Sections of 3 μm in thickness were cut with a microtome from the paraffin-embedded tissue blocks of normal breast tissue, pre-invasive lesions and IBCs (invasive breast cancer). Tissue sections were dewaxed and rehydrated through a series of xylene and ethanol washes. Immunostaining was performed on a Dako automated system. Primary antibody against PKD1 (Cell signaling, Danvers, MA) was incubated overnight at 4°C (dilution 1/100).
Patient-derived xenografts were fixed in 10% neutral buffered formalin and embedded in paraffin. Tissue sections were immunostained in a Discovery XT Platform (Ventana Medical System, Tucson, Arizona, USA, part of Roche Diagnostics) using EDTA buffer pH 8.0 (CC1, Ventana Medical System) for antigen retrieval. Primary antibody against PKD1 (Cell signaling, Danvers, MA) was incubated during 30 min at 37°C (dilution 1/100). After incubation with anti-rabbit secondary antibodies, slides were covered with the chromogenic substrate diaminobenzidine (ChromoMap Kit with Anti rabbit OmniMap, Ventana Medical System) and counterstained with hematoxylin.

Spasojevic C., Marangoni E., Vacher S., Assayag F., Meseure D., Château-Joubert S., Humbert M., Karam M., Ricort J.M., Auclair C., Regairaz M, & Bièche I. (2018). PKD1 is a potential biomarker and therapeutic target in triple-negative breast cancer. Oncotarget, 9(33), 23208-23219.

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Immunohistochemistry of Colon Tissue

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Paraffin-embedded 5 μM sections of distal colon were deparaffinized, rehydrated through graded ethanol, and morphologically checked with H&E. For immunohistochemistry, sections were stained with the following antibodies: monoclonal mouse anti-human cytokeratin 20 (CK20, clone Ks20.8, M7019, Dako, 1:500) and monoclonal mouse anti-E-Cadherin (clone 36, 790-4497, Ventana). All immunostains were performed using Ventana Discovery XT Platform, using CC1 standard regime for CK20 and CC1 mild regime for E-cadherin. Primary antibodies were applied for 60 min followed by biotinylated anti-mouse secondary antibody also for 60 min (Vector, 1:200). Visualization was performed using Ventana DAB map kit. Quantitation of E-cadherin⁺ and CK20⁺ cells was done using Fiji software [13 (link)].

Martin M.L., Zeng Z., Adileh M., Jacobo A., Li C., Vakiani E., Hua G., Zhang L., Haimovitz-Friedman A., Fuks Z., Kolesnick R, & Paty P.B. (2017). Logarithmic expansion of LGR5+ cells in human colorectal cancer. Cellular signalling, 42, 97-105.

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Immunohistochemical analysis of RB1 and SLFN11

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Xenografted tumors were fixed in 10% neutral buffered formalin, embedded in paraffin, and stained with hematoxylin and eosin. Immunostaining was performed on a DISCOVERY XT Platform (Ventana Medical Systems, part of Roche Diagnostics). The slides were incubated with a monoclonal mouse antibody against RB1 (no. 9309, clone 4H1, Cell Signaling Technology) and a polyclonal rabbit antibody against SLFN11 (no. HPA023030, Sigma-Adrich). Slides immunostained with mouse and rabbit immunoglobulin G (IgG) were used as negative controls. Slides were incubated with anti-rabbit/mouse secondary antibodies (horseradish peroxidase complex) and DAB (3,3′-diaminobenzidine tetrahydrochloride) as the substrate for color development (ChromoMap Kit with anti-rabbit OmniMap, Ventana Medical Systems). Immunostaining of RB1 was performed as detailed in previous works (14 (link), 28 (link)). Expression of SLFN11 was quantified with the H-score: Sections were scored for intensity (0 to 3+) and extent (0 to 100%) of staining. By multiplying intensity and extent of staining, each tumor was assigned an H-score (range: 0 to 300). We considered a tumor SLFN11 negative with H-score = 0, SLFN11 low with an H-score between 1 and 60, and SLFN11 high when the H-score was higher than 60.

Coussy F., El-Botty R., Château-Joubert S., Dahmani A., Montaudon E., Leboucher S., Morisset L., Painsec P., Sourd L., Huguet L., Nemati F., Servely J.L., Larcher T., Vacher S., Briaux A., Reyes C., La Rosa P., Lucotte G., Popova T., Foidart P., Sounni N.E., Noel A., Decaudin D., Fuhrmann L., Salomon A., Reyal F., Mueller C., Brugge P.T., Jonkers J., Poupon M.F., Stern M.H., Bièche I., Pommier Y, & Marangoni E. (2020). BRCAness, SLFN11, and RB1 loss predict response to topoisomerase I inhibitors in triple-negative breast cancers. Science translational medicine, 12(531), eaax2625.

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Quantifying Immune Cell Populations in FFPE Tissue

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FFPE specimens were sectioned at five-micrometers and stained on the DISCOVERY XT platform (Ventana Medical Systems, Inc., Tucson, U.S.A). TAM and microglia were identified using antibodies against IBA1. Antibodies targeting CD3 and CD8 stained the entire T cell population and cytotoxic T cells, respectively. Regulatory T cells were stained by antibodies against forkhead box P3 (FOXP3). Due to the lack of a validated antibody, the CD4⁺ T cell number was estimated to be the difference between the CD3⁺ and CD8⁺ T cell numbers according to published protocol.³⁵ In six cases, the number of CD8⁺ T cells exceeded or was identical to the number of CD3⁺ T cells, so that the CD4⁺ T cell population was considered to be equal to the FOXP3⁺ T cell number, since the latter constitutes a subpopulation of CD4⁺ T cells. The following primary antibodies were used: Anti-IBA1 (1:500, rabbit polyclonal, #019–19741, Wako Pure Chemical Ind., Ltd., Osaka, Japan); anti-human FOXP3, clone 259D (1:100, mouse monoclonal, #320202, BioLegend, San Diego, U.S.A); anti-human CD8, clone C8/144B (1:100, mouse monoclonal, code M7103, Dako, Glostrup, Denmark); anti-human CD3 (1:100, rabbit polyclonal, code A0452, Dako, Glostrup, Denmark).

Kaffes I., Szulzewsky F., Chen Z., Herting C.J., Gabanic B., Velázquez Vega J.E., Shelton J., Switchenko J.M., Ross J.L., McSwain L.F., Huse J.T., Westermark B., Nelander S., Forsberg-Nilsson K., Uhrbom L., Maturi N.P., Cimino P.J., Holland E.C., Kettenmann H., Brennan C.W., Brat D.J, & Hambardzumyan D. (2019). Human Mesenchymal glioblastomas are characterized by an increased immune cell presence compared to Proneural and Classical tumors. Oncoimmunology, 8(11), e1655360.

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Immunohistochemical Analysis of Hemoglobin

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Tissue sections (4 µm) were cut with a Leica RM2245 Semi-automated Rotary Microtome and were adhered to Superfrost Plus slides (MICROM, Walldorf, Germany), deparaffinized in xylene and hydrated in a graded series of alcohol. Immunostaining was performed in a Discovery XT Platform (Ventana Medical System, Tucson, Arizona, USA, part of Roche Diagnostics) with antigen retrieval using EDTA buffer, pH 8 (CC1, Ventana Medical System) for primary antibodies anti-HbB (ref OM-B4339, purchased from Omics) and anti-HbA1 (ref GTX42177, purchased from Genetex). Incubation and color development used biotinylated goat anti-mouse secondary antibody, and streptavidin-horseradish peroxidase complex with DAB as substrate (DABMap Kit with Universal Secondary Antibody, Ventana Medical System).

Lerebours F., Vacher S., Guinebretiere J.M., Rondeau S., Caly M., Gentien D., Van Laere S., Bertucci F., de la Grange P., Bièche L., Liang X, & Callens C. (2020). Hemoglobin overexpression and splice signature as new features of inflammatory breast cancer?. Journal of Advanced Research, 28, 77-85.

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Immunohistochemical Analysis of HORMAD1 in Xenografts

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Xenografted tumors were fixed in 10% neutral‐buffered formalin, embedded in paraffin and stained with hematoxylin and eosin. Immunostaining was performed on a Discovery XT Platform (VentanaMedical System, Cambdrige, UK, part of RocheDiagnostics), as previously detailed [15 (link)]. The slides were incubated with a rabbit polyclonal antibody against HORMAD1 (SIGMA, #HPA037850). Slides immunostained with rabbit IgG were used as negative controls. Slides were incubated with an anti‐rabbit secondary antibodies (horseradish peroxidase complex) and 3,30‐diaminobenzidine tetrahydrochloride (DAB) as the substrate for color development (ChromoMap Kit with Anti‐rabbit OmniMap, Ventana Medical System). HORMAD1 immunostaining was assessed by determining the intensity and distribution of stained cancer cells. Expression of HORMAD1 was quantified with the H‐score: Sections were scored for intensity (0–3+) and extent (0–100%) of staining. By multiplying intensity and extent of staining, each tumor was assigned an H‐score (range: 0–300). We considered a tumor HORMAD1 negative with H‐score = 0, HORMAD1 low with an H‐score between 1 and 70, and HORMAD1 high when the H‐score was higher than 70, 70 being the median H‐score.

El‐Botty R., Vacher S., Mainguené J., Briaux A., Ibadioune S., Dahmani A., Montaudon E., Nemati F., Huguet L., Sourd L., Morriset L., Château‐Joubert S., Dubois T., Maire V., Lidereau R., Rapinat A., Gentien D., Coussy F., Bièche I, & Marangoni E. (2023). HORMAD1 overexpression predicts response to anthracycline–cyclophosphamide and survival in triple‐negative breast cancers. Molecular Oncology, 17(10), 2017-2028.

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