After Raman imaging, the tissues were fixed in 4% paraformaldehyde (4 °C overnight) and subsequently processed to be embedded in paraffin. Tissue sections (5 μm) were stained with hematoxylin and eosin (H&E), and immunohistochemistry staining was performed using the DISCOVERY XT biomarker platform (Ventana, Tucson, AZ, USA). Heat-induced epitope retrieval was performed using citrate buffer (pH 6.0). The primary antibodies were diluted as follows: anti-CD44 antibody (1:500; #550538; BD Biosciences, San Jose, CA, USA), anti-Ki-67 antibody (1:250, VP-RM04, Vector Laboratories Inc., Burlingame, CA, USA), anti-CCND1 antibody (1:500; Abcam, Cambridge, MA, USA), and anti-PEG antibody (1:100, PEG-B-47, ab51257, Abcam). All biotin-labeled secondary antibodies, including anti-rabbit antibody (1:300, BA-1000) and anti-rat antibody (1:300, BA-9400), were purchased from Vector Laboratories. All murine specimens were examined by a veterinary pathologist (JRW) from the Tri-Institutional Laboratory of Comparative Pathology who was blinded to the Raman images. All rat specimens were examined by a pathologist (MvdR) from the Department of Pathology at Stanford University who was blinded to the Raman images.
Discovery xt biomarker platform
Manufactured by Roche
Sourced in United States
The Discovery XT biomarker platform is a laboratory equipment product developed by Roche. It is designed to facilitate the detection and analysis of biomarkers, which are molecules that can indicate the presence or progression of a particular disease or condition. The platform provides researchers and clinicians with the tools to perform various assays and tests, enabling them to gather valuable data and insights. The core function of the Discovery XT is to support the identification and measurement of biomarkers, which can contribute to advancements in medical research and diagnostic applications.
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Lab products found in correlation
Ab9610, by Merck Group (2 mentions)
Ba 1000, by Vector Laboratories (2 mentions)
Ba 9400, by Vector Laboratories (1 mentions)
Anti ki 67 antibody, by Vector Laboratories (1 mentions)
Vp rm04, by Vector Laboratories (1 mentions)
Anti ccnd1 antibody, by Abcam (1 mentions)
Peg b 47, by Abcam (1 mentions)
Anti cd44 antibody, by BD (1 mentions)
Clone epr3864, by Abcam (1 mentions)
M0851, by Agilent Technologies (1 mentions)
Ab51257, by Abcam (1 mentions)
Ab76609, by Abcam (1 mentions)
Discovery dab map detection kit, by Roche (1 mentions)
Jbw301, by Merck Group (1 mentions)
Peroxidase histostain plus kit, by Zymo Research (1 mentions)
Hematoxylin, by Roche (1 mentions)
3 3 diaminobenzidine (dab), by Zymo Research (1 mentions)
6 protocols using discovery xt biomarker platform
1
Histological and Immunohistochemical Analysis of Raman Imaging Samples
2
Immunohistochemical Analysis of SPINK1 and ERG
3
Multimodal Imaging of Glioblastoma
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Intact GBM-bearing brains were sliced coronally (1 mm slice thickness) and embedded in paraffin. 5 μm-thick continuous sections were cut for hematoxylin and eosin (H&E) staining and immunohistochemistry (IHC) staining, followed by high-resolution Raman imaging on paraffin blocks. IHC staining was performed on the Discovery XT biomarker platform (Ventana, Tucson, AZ) as previously described 30 . Antibodies for OLIG2 (1:300, AB9610, Millipore, Temecula, CA), polyethylene glycol (1:100, ab51257, Abcam, Cambridge, MA), ITGB3 (1:100, 13166, Cell Signaling, Danvers, MA), ITGAV (1:2000, ab76609, Abcam), HA-tag (1:200, 11867423001, Roche, San Francisco, CA), IBA1 (1:600, 019-19741, Wako, Richmond, VA), NOS3 (1:200, 610296, BD Biosciences, Franklin Lakes, NJ) and ACTA2 (1:350, M0851, DAKO, Carpinteria, CA) were used as the primary antibodies. The slides were digitally scanned with Pannoramic Flash (3DHistech, Hungary) and relevant tissue areas were exported into tiff format. Quantification of Olig2 was performed using ImageJ/FIJI (NIH). Color deconvolution algorithm was used to determine the area of positive signal, which was normalized to tissue area.
4
Immunohistochemical Analysis of Olig2 in Brain Tumor Tissues
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Brain tissues after tumor resections were embedded in paraffin. Hematoxylin and eosin staining and immunohistochemistry (IHC) staining were performed on 5 μm-thick sections. The Discovery XT biomarker platform (Ventana, Tucson, AZ) was used for IHC staining of Olig2. Antigen was retrieved using the heat-induced antigen unmasking technique in citrate-based buffer (pH 6.0). Anti-Olig-2 (1:300, AB9610, Millipore, Temecula, CA) and biotin-labeled anti-rabbit antibody (1:300, BA-1000, Vector Laboratories, Burlingame, CA) were used as the primary and secondary antibodies, respectively. Streptavidin–biotin peroxidase was visualized by the Discovery DAB map detection kit (760-124, Ventana).
5
Immunohistochemical Analysis of SLC6A2/NET
6
Multiplexed Immunohistochemistry for Breast Cancer Markers
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Paraffin-embedded human breast tissue sections and PDX were deparaffinized in xylene and rehydrated in graded alcohol. Immunohistochemistry was performed on a Ventana Discovery XT biomarker platform. For Ki-67 and cleaved-caspase-3 staining, antigen enhancement was performed by incubating the sections in citrate buffer, pH 6 (Dakocytomation). Staining was done using Peroxidase histostain-Plus Kit (Zymed) according to the manufacturer’s protocol. Ki-67 antibody (#R626, RTU, Agilent) and cleaved-caspase 3 (#9661 S, RTU, Cell Signaling) was used. DAB (Zymed) was used as substrate for peroxidase. For the multiplexed immunostaining of ALDH1 and γH2AX, we start with the anti-ALDH1 (mAb clone 44, Becton Dickinson, 1/50) antibody followed with the anti-γH2AX antibody (mAb JBW301, Merck Millipore, 1/2000). Multiplex staining was performed using Discovery OmniMap Multimer HRP with DAB substrate (Ventana) for ALDH1 and Discovery Purple kit substrate (Ventana) for γH2AX. Slides were counterstained with hematoxylin (Ventana).
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