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Bond platform

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The Bond platform is a versatile laboratory instrument designed for high-throughput sample processing and analysis. It features automated liquid handling capabilities, enabling efficient and precise sample preparation and assay workflows.

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Lab products found in correlation

Benchmark ultra, by Leica (2 mentions) Anti human cd8, by Cell Marque (2 mentions) Anti human pd l1, by Agilent Technologies (2 mentions) Rnascope probe, by Advanced Cell Diagnostics (1 mentions) Dako immunostainer, by Agilent Technologies (1 mentions) Buffer w, by NanoString (1 mentions) Ab196147, by Novus Biologicals (1 mentions) Mabn92 af647, by Abcam (1 mentions) Nbp2 33184af532, by Thermo Fisher Scientific (1 mentions) S7575, by Thermo Fisher Scientific (1 mentions) Axio scan z1 slide scanner, by Zeiss (1 mentions) Prolong antifade mounting medium, by Thermo Fisher Scientific (1 mentions) Tissue tek vip 6, by Sakura Finetek (1 mentions) Histocore arcadia embedding center, by Leica (1 mentions) Opal multiplex ihc kit, by Akoya Biosciences (1 mentions) Ar1068, by Boster Bio (1 mentions) Halo image analysis software, by Indica Labs (1 mentions) Rhodamine red x conjugated goat anti rabbit igg, by Jackson ImmunoResearch (1 mentions) Smad4, by Santa Cruz Biotechnology (1 mentions) Clone c4, by Santa Cruz Biotechnology (1 mentions)

Close spelling variants of this product

BOND Rx platform (24 citations) Bond-III platform (12 citations) Bond Max platform (3 citations) BOND-RXm platform (3 citations) Bond RX Autostainer platform (2 citations) Bond automated staining platform (2 citations) Bond staining platform (2 citations) Leica Bond IHC platform (2 citations) Leica-Bond stainer platform (2 citations) BOND Rx automated staining platform (2 citations)

16 protocols using bond platform

1

Bulk RNAseq and Single-cell Analysis of Retinal Transcriptome

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Bulk RNAseq of retinas and differentially expressed gene analysis was performed as previously described in (62 (link)). For bulk RNAseq transcriptomic studies, the total RNA was prepared from the retinas and livers of five Nfe2l1OE (four males and one female) and WT (four males and one female) littermate mice of 6 to 7 weeks of age. Differentially expressed genes are shown in Microsoft Excel files data S1 (retina) and S2 (liver). RT-qPCR was performed using primers listed in table S4 as previously described (24 (link)). RNA ISH was performed on 5-μm–thick paraffin sections prepared from formalin-fixed eyes with RNAscope probes (Advanced Cell Diagnostics, Hayward, CA, USA) listed in table S5 on the automated Leica Bond platform (Leica Microsystems GmbH, Wetzlar, Germany) following the manufacturer’s instructions. The single-cell libraries were prepared with pooled retinas from one male and one female littermate mice (Nfe2l1OE and WT) using 10x Chromium Platform and following protocols described in (63 (link)). Sequencing was performed at UF Interdisciplinary Center for Biotechnology Research, and single-cell data analysis was performed as described in our previous studies (62 (link), 64 (link)).
Wang Y., Snell A., Dyka F.M., Colvin E.R., Ildefonso C., Ash J.D, & Lobanova E.S. (2023). Overexpression of Nfe2l1 increases proteasome activity and delays vision loss in a preclinical model of human blindness. Science Advances, 9(28), eadd5479.
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2

Standardized Immunohistochemical Profiling

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Five-micron tissue sections were cut from the selected block. Immunostaining assays were performed on a Leica Bond Platform (Leica Microsystems, Chicago, IL, USA) for French cases and an automated DAKO Immunostainer for Italian cases.
The antibodies were used according to the manufacturer's protocol. The same panel of antibodies was applied to all cases. The antibodies/antisera used are listed in Table 1. A stain was considered 'positive' when at least 5% of neoplastic cells showed immunoreactivity in the appropriate site.
Denize T., Massa S., Valent A., Militti L., Bertolotti A., Barisella M., Rioux-Leclercq N., Malouf G.G., Spreafico F., Verschuur A., van der Beek J., Tytgat L., van den Heuvel-Eibrink M.M., Vujanic G., Collini P, & Coulomb A. (2022). Renal cell carcinoma in children and adolescents: a retrospective study of a French-Italian series of 93 cases. Histopathology, 80(6).
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3

Immunohistochemical Detection of LFABP in Tumors

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Immunohistochemical staining for LFABP (rabbit polyclonal antibody, 1:50 dilution; Abcam, Cambridge, United Kingdom) was performed on formalin-fixed, paraffin-embedded tissue using standard methodology on the Leica BOND platform (Leica Biosystems, Nussloch, Germany), with appropriate positive and negative controls. Only complete loss of staining for LFABP throughout the tumor was considered negative (Table 2, “−”); all other staining intensities and patterns were considered positive (Table 2, “+”).
Cho S.J., Ferrell L.D, & Gill R.M. (2015). Expression of liver fatty acid binding protein in hepatocellular carcinoma. Human pathology, 50, 135-139.
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4

GeoMx DSP Slide Preparation Protocol

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Slides were prepared by baking in a drying oven and then processed using the Leica BOND platform (Leica Biosystems) as specified by the NanoString GeoMx DSP Slide Preparation User Manual. Briefly, slides were deparaffinized with xylene then rehydrated through a graded ethanol series. Targets were exposed by incubating the slides in a pressure cooker at 100°C in 1X Tris-EDTA buffer, pH9 followed by proteinase K digestion. Next, the GeoMx NGS-RNA Probe Mix (NanoString Technologies) was applied to each slide, and slides were incubated at 37°C for 16-24 hours. This probe mix contained barcoded oligonucleotide probes against 1,700 gene targets and included internal positive and negative control probes. After washing, the slides were then blocked in Buffer W (NanoString Technologies) and stained with morphology markers for 1 hour. The fluorescently conjugated morphology markers used are as follows: Iba1 at 1:75 dilution (EMD Millipore, MABN92-AF647), CD3 at 1:50 dilution (Abcam, ab196147), GFAP at 1:3000 dilution (Novus Biologicals, NBP2-33184AF532), and SYTO 13 nuclei acid stain at 1:5 dilution (Thermo Scientific S7575).
Kim Y., Danaher P., Cimino P.J., Hurth K., Warren S., Glod J., Beechem J.M., Zada G, & McEachron T.A. (2023). Highly multiplexed spatially resolved proteomic and transcriptional profiling of the glioblastoma microenvironment using archived formalin-fixed paraffin-embedded specimens.. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc, 36(1), 100034.
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5

Histological Analysis of Enucleated Eyeballs

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Eyes were enucleated and fixed in 4% paraformaldehyde (AR1068; Boster Bio, Pleasanton, CA, USA) for 48 hours at 4°C. Whole eyeballs were dehydrated and infiltrated with paraffin using a tissue processor (Tissue-Tek VIP 6; Sakura Finetek USA, Inc., Torrance, CA, USA) and subsequently embedded into paraffin blocks on a HistoCore Arcadia Embedding Center (Leica Biosystems, Melbourne, Australia). Three-micrometer sections were cut at the level of the optical nerve and hematoxylin and eosin and Masson's Trichrome staining was performed according to standard protocols. Immunofluorescence staining was carried out on the automated Leica Bond platform (Leica Biosystems) using the Opal Multiplex IHC Kit (Akoya Biosciences, Menlo Park, CA, USA). Antibodies used in this study and details about the antigen retrieval are listed in the Table. Nuclei were stained with spectral DAPI (FP1490; Akoya Biosciences) and slides mounted with ProLong Antifade Mounting Medium (P36961; Invitrogen, Carlsbad, CA, USA). Tissues were imaged using an Axio Scan.Z1 slide scanner (magnification ×20; Carl Zeiss Microscopy GmbH, Jena, Germany). Iba1+ and F4/80+ area was quantified using HALO image analysis software (Indica Labs, Albuquerque, NM, USA).
Weigelt C.M., Zippel N., Fuchs H., Rimpelä A.K., Schönberger T., Stierstorfer B., Bakker R.A, & Redemann N.H. (2022). Characterization and Validation of In Vitro and In Vivo Models to Investigate TNF-α-Induced Inflammation in Retinal Diseases. Translational Vision Science & Technology, 11(5), 18.
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6

Immuno-Histochemical Tissue Analysis

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For H&E staining, formalin-fixed, paraffin-embedded tissue cut at 4 microns and stained using the Tissue-Tek Prisma automated slide stainer. Immunostaining was performed on the Ventana Benchmark Ultra platform (CD3) and the Leica Bond platform (CD8, PD-L1). Antibodies used include anti-human CD3 (cat. no. 103A-76, Cell Marque), anti-human CD8 (cat. no. M7103, Dako) and anti-human PD-L1 (cat. no. 13684S, Cell Signaling Technology).
Yost K.E., Satpathy A.T., Wells D.K., Qi Y., Wang C., Kageyama R., McNamara K., Granja J.M., Sarin K.Y., Brown R.A., Gupta R.K., Curtis C., Bucktrout S.L., Davis M.M., Chang A.L, & Chang H.Y. (2019). Clonal replacement of tumor-specific T cells following PD-1 blockade. Nature medicine, 25(8), 1251-1259.
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7

Immuno-Histochemical Tissue Analysis

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For H&E staining, formalin-fixed, paraffin-embedded tissue cut at 4 microns and stained using the Tissue-Tek Prisma automated slide stainer. Immunostaining was performed on the Ventana Benchmark Ultra platform (CD3) and the Leica Bond platform (CD8, PD-L1). Antibodies used include anti-human CD3 (cat. no. 103A-76, Cell Marque), anti-human CD8 (cat. no. M7103, Dako) and anti-human PD-L1 (cat. no. 13684S, Cell Signaling Technology).
Yost K.E., Satpathy A.T., Wells D.K., Qi Y., Wang C., Kageyama R., McNamara K., Granja J.M., Sarin K.Y., Brown R.A., Gupta R.K., Curtis C., Bucktrout S.L., Davis M.M., Chang A.L, & Chang H.Y. (2019). Clonal replacement of tumor-specific T cells following PD-1 blockade. Nature medicine, 25(8), 1251-1259.
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8

NELL1 Immunohistochemistry in Malignancy

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Three tumor samples from two patients, including one biopsy and mastectomy specimen from a patient with invasive ductal carcinoma of the breast, and one lymph node resection from a patient with follicular lymphoma were stained with rabbit polyclonal anti-NELL1 antibody. Both patients had concurrent membranous nephropathy at the time of an active malignancy. Immunohistochemical staining for NELL1 was performed on 4 μm sections of tumor tissue on the Leica BOND platform at 1:100 primary dilution with anti-NELL1 rabbit polyclonal antibody (Thermo Fisher Scientific, PA5–27958). Rhodamine red X-conjugated goat anti-rabbit IgG (1:100; Jackson Immunoresearch, cat# 111–295-144) was used as the secondary antibody.
Caza T., Hassen S., Dvanajscak Z., Kuperman M., Edmondson R., Herzog C., Storey A., Arthur J., Cossey L.N., Sharma S., Kenan D, & Larsen C. (2020). NELL1 is a target antigen in malignancy-associated membranous nephropathy.. Kidney international, 99(4), 967-976.
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9

Immunohistochemical Analysis of Pancreatic Tumors

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Immunohistochemistry of paraffin-embedded primary pancreatic tumor tissue was performed by the immunohistochemistry laboratory at the Johns Hopkins Hospital. In brief, paraffin was removed from a sample before cell conditioning and incubation. Commercially available antibodies to p53 (Ventana) and SMAD4 (Santa Cruz Biotechnology) were individually applied to separate samples and allowed to further incubate, before removal and counter-staining with hematoxylin. A Ventana BenchMark ULTRA platform was used to process the p53 slides, whereas a Leica BOND platform was used for SMAD4. A pathologist specializing in pancreatic tumors reviewed all immunolabeled slides to determine the presence or absence of SMAD4 and p53, in addition to whether labeling was normal or abnormal as has been described.40 (link)
Poruk K.E., Valero V I.I.I., Saunders T., Blackford A.L., Griffin J.F., Poling J., Hruban R.H., Anders R.A., Herman J., Zheng L., Rasheed Z.A., Laheru D.A., Ahuja N., Weiss M.J., Cameron J.L., Goggins M., Iacobuzio-Donahue C.A., Wood L.D, & Wolfgang C.L. (2016). Circulating Tumor Cell Phenotype Predicts Recurrence and Survival in Pancreatic Adenocarcinoma. Annals of surgery, 264(6), 1073-1081.
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10

Comprehensive IHC Protocol for BAP1, p16, MTAP

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Standardized protocol using the DAKO platform was used for IHC on FFPE tissue for BAP1 (1:50 primary, Clone C4; Santa Cruz Biotech, TX), p16 (1:400 primary, Clone JC2; Cell Marque, CA), and MTAP (1:100 primary, Clone 2G4; Abnova, Taipei, Taiwan). IHC for H3 K27me3 (1:600 primary, Clone C36B11; Cell Signaling, MA) was performed on the Leica BOND platform.
Tosefsky K., Martin K.C., Rebchuk A.D., Wang J.Z., Nassiri F., Lum A., Zadeh G., Makarenko S, & Yip S. (2024). Molecular prognostication in grade 3 meningiomas and p16/MTAP immunohistochemistry for predicting CDKN2A/B status. Neuro-Oncology Advances, 6(1), vdae002.
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