Discovery xt autostainer

Manufactured by Roche

Sourced in United States

The Discovery XT Autostainer is a laboratory instrument designed for automated slide staining. It is capable of performing immunohistochemistry (IHC) and in situ hybridization (ISH) staining protocols, enabling efficient and consistent processing of tissue samples.

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Lab products found in correlation

Superfrost plus slide, by Thermo Fisher Scientific (7 mentions) Aperio at turbo scanner, by Leica (4 mentions) Permount, by Thermo Fisher Scientific (3 mentions) Aperio imagescope software, by Leica (2 mentions) Omnimap rabbit polyclonal hrp, by Roche (2 mentions) Chromomap dab, by Roche (2 mentions) Ventana discovery ultra instrument, by Roche (2 mentions) Cv5030 coverslipper, by Leica (2 mentions) Clone ln10, by Leica (1 mentions) Aperio scanscope system, by Leica (1 mentions) Phospho smad1 smad5 smad8 antibody, by Cell Signaling Technology (1 mentions) 3 3 diaminobenzidine dab peroxidase substrate kit, by Vector Laboratories (1 mentions) Ndp view software, by Hamamatsu Photonics (1 mentions) Nanozoomer ht, by Hamamatsu Photonics (1 mentions) Af 307 na, by R&D Systems (1 mentions) P53 clone bp53 11, by Roche (1 mentions) Alexa fluor 488 goat anti mouse igg, by Thermo Fisher Scientific (1 mentions) Aperio imagescope, by Leica (1 mentions) Ultraview sish detection kit, by Roche (1 mentions)

Spelling variants of this product

Discovery XT (190 citations) Autostainer (13 citations) Ventana Discovery XT autostainer (7 citations) Ventana Discovery XT Autostainer platform (4 citations) Discovery Ultra XT autostainer (3 citations)

21 protocols using discovery xt autostainer

Immunohistochemical Staining of PTHLH, RUNX2, and Ki-67

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PTHLH, RUNX2 and Ki-67 IHC staining was performed using an automated immunostainer (Ventana Discovery XT autostainer, Ventana, USA). Antigens were retrieved by heat-induced antigen retrieval for 30 minutes with TRIS-EDTA buffer. Slides were stained with polyclonal rabbit PTHLH antibody (1:200; GeneTex, Taiwan), monoclonal mouse RUNX2 antibody (1:20; Santa Cruz Biotechnology, CA), and monoclonal mouse Ki-67 antibody (1:100; Dako, Denmark).
For PTHLH IHC staining interpretation, both the immunoreactivity intensity and percentage were recorded. The intensity of staining was defined as 0, no staining; 1+, weak staining; 2+, moderate staining; 3+, strong staining. The extent of staining was scored by the percentage of positive cells (0–100%). The final IHC scores (0–300) were the results of staining intensity score multiplied by the percentage of positive cells. Then, all cases were divided into two groups according to the final IHC scores. A score more than and include 150 itself was defined as high IHC expression level and a score less than 150 was defined as low expression.

Chang W.M., Lin Y.F., Su C.Y., Peng H.Y., Chang Y.C., Hsiao J.R., Chen C.L., Chang J.Y., Shieh Y.S., Hsiao M, & Shiah S.G. (2017). Parathyroid Hormone-Like Hormone is a Poor Prognosis Marker of Head and Neck Cancer and Promotes Cell Growth via RUNX2 Regulation. Scientific Reports, 7, 41131.

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Multiplex Immunofluorescence Profiling of FFPE Tumor Sections

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FFPE tumor sections were profiled by multiplex immunofluorescence using a Discovery XT autostainer (Ventana Medical Systems) with appropriate controls. Antibodies used were CD3 (Leica, Clone LN10, 1:100 dilution), CD20 (Leica, Clone L26, 1:200 dilution) and Iba1 (Wako Chemicals, 019-19741, 1:500 dilution).

Wang L., Jung J., Babikir H., Shamardani K., Jain S., Feng X., Gupta N., Rosi S., Chang S., Raleigh D., Solomon D., Phillips J.J, & Diaz A.A. (2022). A single-cell atlas of glioblastoma evolution under therapy reveals cell-intrinsic and cell-extrinsic therapeutic targets. Nature Cancer, 3(12), 1534-1552.

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Immunohistochemical Analysis of Patient Samples

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Immunohistochemistry (IHC) for patient samples and patient-derived xenografts was performed at the Segal Cancer Centre Research Pathology Facility (Jewish General Hospital). The slides were stained using the Discovery XT Autostainer (Ventana Medical System). All solutions used for automated immunohistochemistry were from Ventana Medical System (Roche) unless otherwise specified. After de-paraffinization and heat-induced epitope retrieval (CC1 prediluted solution Ref: 950–124, standard protocol), sections were incubated with primary antibodies: anti-p-Smad1/5/9 (clone D5B10, CST 13820) and anti-NKX6.1 (clone EPR20405, abcam) diluted at 1:100 and 1:250, respectively. Slides were counterstained with hematoxylin and Bluing Reagent, and finalized with mounting medium (Eukitt, Fluka Analytical). Sections were scanned using the Aperio AT Turbo Scanner (Leica Biosystems).

Jessa S., Mohammadnia A., Harutyunyan A.S., Hulswit M., Varadharajan S., Lakkis H., Kabir N., Bashardanesh Z., Hébert S., Faury D., Vladoiu M.C., Worme S., Coutelier M., Krug B., Andrade A.F., Pathania M., Bajic A., Weil A.G., Ellezam B., Atkinson J., Dudley R.W., Farmer J.P., Perreault S., Garcia B.A., Larouche V., Blanchette M., Garzia L., Bhaduri A., Ligon K.L., Bandopadhayay P., Taylor M.D., Mack S.C., Jabado N, & Kleinman C.L. (2022). K27M in canonical and noncanonical H3 variants occurs in distinct oligodendroglial cell lineages in brain midline gliomas. Nature genetics, 54(12), 1865-1880.

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Automated Immunohistochemistry of Ferroportin

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Tissue specimens from 3 animals per experimental group were fixed in 10% buffered formalin and embedded in paraffin. One section per animal was prepared for liver and spleen specimens, and three sections per animal for duodenal specimens. Samples were cut at 4 µm, placed on SuperFrost/Plus slides (Thermo Fisher Scientific, Waltham, MA), and dried overnight at 37°C. The slides were then loaded onto the Discovery XT Autostainer (Ventana Medical Systems, Oro Valley, AZ) and underwent de‐paraffinization and heat‐induced epitope retrieval for automated immunohistochemistry. Immunostaining was performed by using a 1:500 diluted custom‐made ferroportin antibody⁽¹⁶⁾ and the Omnimap rabbit polyclonal horseradish peroxidase (#760‐4311) detection kit.⁽¹⁷⁾ Slides were counterstained with hematoxylin, blued with Bluing Reagent, washed in warm soapy water, dehydrated through graded alcohols, cleared in xylene, and mounted with Permount (Thermo Fisher Scientific). Immunostained tissue slides were scanned using the Aperio ScanScope System (Leica BioSystems [Aperio], Vista, CA) at ×20 or ×40 magnifications, to provide high‐resolution digital images. Slides with hematoxylin‐stained splenic sections were analyzed and quantified for red and white pulp content by using the Aperio ImageScope software (Leica Biosystems).

Katsarou A., Gkouvatsos K., Fillebeen C, & Pantopoulos K. (2021). Tissue‐Specific Regulation of Ferroportin in Wild‐Type and Hjv‐/‐ Mice Following Dietary Iron Manipulations. Hepatology Communications, 5(12), 2139-2150.

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Immunohistochemistry of Ferroportin in Mice

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Tissue specimens were fixed in 10% buffered formalin and embedded in paraffin. Samples from three different mice for each experimental condition were cut at 4 µm, placed on SuperFrost/Plus slides (Fisher) and dried overnight at 37°C. The slides were then loaded onto the Discovery XT Autostainer (Ventana Medical System) for automated immunohistochemistry. Slides underwent deparaffinization and heat-induced epitope retrieval. Immunostaining was performed by using 1:500 diluted rabbit polyclonal antibodies against ferroportin (Maffettone et al., 2010 (link)) and an appropriate detection kit (Omnimap rabbit polyclonal HRP, #760–4311 and ChromoMap-DAB #760–159; Roche). Negative controls were performed by the omission of the primary antibody. Slides were counterstained with hematoxylin for 4 min, blued with Bluing Reagent for 4 min, removed from the autostainer, washed in warm soapy water, dehydrated through graded alcohols, cleared in xylene, and mounted with Permount (Fisher). Sections were analyzed by conventional light microscopy and quantified by using the Aperio ImageScope software (Leica Biosystems; Fillebeen et al., 2018 (link)).

Charlebois E., Fillebeen C., Katsarou A., Rabinovich A., Wisniewski K., Venkataramani V., Michalke B., Velentza A, & Pantopoulos K. (2022). A crosstalk between hepcidin and IRE/IRP pathways controls ferroportin expression and determines serum iron levels in mice. eLife, 11, e81332.

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Immunohistochemical Analysis of BMP4 Signaling

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Representative three 1-mm-diameter cores from each tumor taken from the formalin-fixed paraffin embedded tissues were selected by morphology typical of the diagnosis. Immunohistochemical (IHC) staining was performed on serial 5-micrometer-thick tissue sections cut from the tissue microarray (TMA). IHC staining of FSTL1, BMP4, and Smad4 was performed using an automated immunostainer (Ventana Discovery XT autostainer, Ventana, USA). The antigens were retrieved by heat-induced antigen retrieval for 30 minutes with TRIS-EDTA buffer. The slides were stained with a polyclonal rabbit FSTL1 antibody (1:250; GeneTex, Taiwan), a polyclonal rabbit BMP4 antibody (1:300; GeneTex, Taiwan), and a polyclonal rabbit Smad4 antibody (1:500; Proteintech, USA). For phospho-Smad1/Smad5/Smad8 (p-Smad1/5/8), manual IHC staining was performed. Briefly, the slides were submitted to heat-induced antigen retrieval for 10 minutes with DAKO antigen retrieval buffer (pH 6) and incubated at 4 °C overnight with a polyclonal rabbit phospho-Smad1/Smad5/Smad8 antibody (1:50; Cell Signaling, USA) and then visualised using the 3, 3′-diaminobenzidine (DAB) peroxidase substrate kit (Vector Laboratories, USA).

Chiou J., Su C.Y., Jan Y.H., Yang C.J., Huang M.S., Yu Y.L, & Hsiao M. (2017). Decrease of FSTL1-BMP4-Smad signaling predicts poor prognosis in lung adenocarcinoma but not in squamous cell carcinoma. Scientific Reports, 7, 9830.

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Immunohistochemical Analysis of Ferroportin and Hepcidin

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Tissue specimens were fixed in 10% buffered formalin and embedded in paraffin. Samples were cut at 4 µm, placed on SuperFrost/Plus slides (Fisher Scientific, Ottawa, ON, Canada), and dried overnight at 37 °C. The slides were then loaded onto the Discovery XT Autostainer (Ventana Medical System, Oro Valley, AZ, USA) for automated immunohistochemistry. Slides underwent deparaffinization and heat-induced epitope retrieval. Immunostaining was performed by using rabbit polyclonal antibodies against ferroportin ([43 (link)]; 1:500 diluted) or hepcidin (Abcam, Cambridge, UK; # ab30760; 1:25 diluted) and an appropriate detection kit (Omnimap rabbit polyclonal HRP, #760-4311 and ChromoMapDAB #760-159; Roche Diagnostics, Mississauga, ON, Canada). Negative controls were performed by the omission of the primary antibody. Slides were counterstained with hematoxylin for four minutes, blued with Bluing Reagent for four minutes, removed from the autostainer, washed in warm soapy water, dehydrated through graded alcohols, cleared in xylene, and mounted with Permount (Fisher Scientific, Ottawa, ON, Canada).

Bigorra Mir M., Charlebois E., Tsyplenkova S., Fillebeen C, & Pantopoulos K. (2023). Cardiac Hamp mRNA Is Predominantly Expressed in the Right Atrium and Does Not Respond to Iron. International Journal of Molecular Sciences, 24(6), 5163.

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Immunohistochemical Analysis of AK4 and AK1

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Tissue cores from each specimen picked according to original hematoxylin and eosin slides and diagnostic information. IHC staining was performed using an automatic immunostainer (Discovery XT autostainer, Ventana, USA). Paraffin sections were dewaxed in a 60 °C oven, deparaffinized in xylene, and then rehydrated in graded EtOH. Antigen retrieval was performed using heat-induced TRIS-EDTA buffer for 30 minutes. Immunoreactivity of protein expressions was developed using 3, 3′-diaminobenzidine (DAB) peroxidase substrate kit (Ventana, USA). The slides were counterstained with hematoxylin. The antibodies were used to determine AK4 and AK1 expression: AK4 (Genetex, 1:200), and AK1 (Genetex, 1:100).

Jan Y.H., Lai T.C., Yang C.J., Huang M.S, & Hsiao M. (2019). A co-expressed gene status of adenylate kinase 1/4 reveals prognostic gene signature associated with prognosis and sensitivity to EGFR targeted therapy in lung adenocarcinoma. Scientific Reports, 9, 12329.

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Immunohistochemistry of H3.3 Mutations

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Immunohistochemistry (IHC) was performed at Histology Platform (RI-MUHC) and the Segal Cancer Centre Research Pathology Facility (Jewish General Hospital). PDOX and mouse brain tissue samples were cut at 4-6 μm, placed on SuperFrost/Plus slides (Fisher) and dried overnight at 37°C, before IHC processing. After de-paraffinization and epitope retrieval, sections were incubated with primary antibodies: Gsx2 (ABN162, Millipore Sigma RRID:AB_11203296) in 1:100; H3.3 G34R (RM240 , RevMAb Biosciences, RRID:AB_2716433) in 1:50; and H3.3 G34V (RM307, RevMAb Biosciences RRID:AB_2716435) in 1:40. Slides were then loaded onto the Discovery XT Autostainer or Ventana Discovery Ultra Instrument (Ventana Medical Systems). Slides were counterstained with hematoxylin, blued with Bluing Reagent, washed, dehydrated through graded alcohols, cleared in xylene, and mounted with mounting medium (Eukitt, Fluka Analytical) or Leica CV 5030 coverslipper. Sections were analyzed by conventional light microscopy or scanned using the Aperio AT Turbo Scanner (Leica Biosystems).

Chen C.C., Deshmukh S., Jessa S., Hadjadj D., Lisi V., Andrade A.F., Faury D., Jawhar W., Dali R., Suzuki H., Pathania M., A D., Dubois F., Woodward E., Hébert S., Coutelier M., Karamchandani J., Albrecht S., Brandner S., De Jay N., Gayden T., Bajic A., Harutyunyan A.S., Marchione D.M., Mikael L.G., Juretic N., Zeinieh M., Russo C., Maestro N., Bassenden A.V., Hauser P., Virga J., Bognar L., Klekner A., Zapotocky M., Vicha A., Krskova L., Vanova K., Zamecnik J., Sumerauer D., Ekert P.G., Ziegler D.S., Ellezam B., Filbin M.G., Blanchette M., Hansford J.R., Khuong-Quang D.A., Berghuis A.M., Weil A.G., Garcia B.A., Garzia L., Mack S.C., Beroukhim R., Ligon K.L., Taylor M.D., Bandopadhayay P., Kramm C., Pfister S.M., Korshunov A., Sturm D., Jones D.T., Salomoni P., Kleinman C.L, & Jabado N. (2020). Histone H3.3G34-mutant interneuron progenitors co-opt PDGFRA for gliomagenesis. Cell, 183(6), 1617-1633.e22.

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Immunohistochemical Detection of LAMP-2 for Phospholipidosis Identification

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To differentiate PL from other changes in the liver, immunostaining for LAMP-2 was conducted as previously described [46 (link)]. LAMP-2 is a promising marker for PL which has been characterized by immunohistochemistry in rat tissues treated with PL-inducing drugs [47 (link)], or cells treated with chloroquine [48 (link)]. Briefly, both hepatocyte culture systems were incubated with 10 μM amikacin, amiodarone, sertraline and 25 μM acetaminophen for 48 h. Paraffin-embedded cell blocks were subsequently made, and each was sectioned and stained automatically using a Discovery XT Autostainer (Ventana Medical Systems). A primary anti-LAMP-2 antibody (Invitrogen) was used at a 1:200 dilution for 40 min. The target antigen was visualized via a DAB reaction as described previously.

Lee J.Y., Han H.J., Lee S.J., Cho E.H., Lee H.B., Seok J.H., Lim H.S, & Son W.C. (2020). Use of 3D Human Liver Organoids to Predict Drug-Induced Phospholipidosis. International Journal of Molecular Sciences, 21(8), 2982.

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