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Leica bond

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The Leica Bond is a specialized laboratory equipment designed for advanced imaging and analysis. It provides high-quality, reliable performance for scientific and research applications. The core function of the Leica Bond is to facilitate comprehensive sample preparation and staining processes, enabling detailed examination and evaluation of various materials and specimens.

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Lab products found in correlation

Fisher chemical permount mounting medium, by Thermo Fisher Scientific (1 mentions) Bond polymer refine detection, by Leica Biosystems (1 mentions) Signalstain antibody diluent, by Cell Signaling Technology (1 mentions) Halo platform, by Indica Labs (1 mentions) Ventana benchmark ultra, by Roche (1 mentions) Clone e6h4, by Roche (1 mentions) Clone g3 245, by BD (1 mentions) Ab178400, by Abcam (1 mentions) Leica bond detection kit, by Leica camera (1 mentions)

6 protocols using leica bond

1

Immunohistochemical Analysis of β-catenin

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FFPE slides were treated sequentially following the Leica BOND RX default Bake & Dewax Protocol, HIER (ER2, 20 min, 95°C) protocol and the immunohistochemistry (IHC) Leica BOND protocol (anti-β-catenin antibody, Cell Signaling Technology, clone: 15B8, cat. 37 447S; SignalStain Antibody Diluent, Cell Signaling Technology, cat. 8112; BOND Polymer Refine Detection, Leica Biosystems, cat. DS9800). Stained slides were treated with 95% ethanol for 10 s twice and 100% ethanol for 10 s twice, followed by xylene for 10 s twice. Slides were mounted with Fisher Chemical Permount Mounting Medium (Fisher Scientific, cat. SP15-100) and scanned on HAMAMATSU NanoZoomer 2.0RS. Images were analyzed using the HALO platform (Indica Labs).
Wong-Rolle A., Dong Q., Zhu Y., Divakar P., Hor J.L., Kedei N., Wong M., Tillo D., Conner E.A., Rajan A., Schrump D.S., Jin C., Germain R.N, & Zhao C. (2022). Spatial meta-transcriptomics reveal associations of intratumor bacteria burden with lung cancer cells showing a distinct oncogenic signature. Journal for Immunotherapy of Cancer, 10(7), e004698.
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2

Immunohistochemical Analysis of Cell Markers

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Immunohistochemical studies were performed on 4-μm formalin-fixed paraffin-embedded sections with commercially available antibodies targeting the following proteins: Rb (1:100 dilution, clone G3–245; BD Biosciences, San Jose, CA, USA), MDM2 (1:200 dilution, clone IF2; MilliporeSigma, Burlington, MA, USA), p16 (1:4 dilution, clone E6H4; Ventana Medical Systems, Roche, Indianapolis, IL, USA), p53 (prediluted, clone DO-7; Ventana Medical Systems), and CD34 (1:160 dilution, clone MY10; BD Biosciences). Immunostaining for Rb, p16 and CD34 was performed on the Leica Bond (Leica, Buffalo Grove, IL, USA), and immunostaining for p16 and p53 was performed on the Ventana BenchMark ULTRA (Ventana Medical Systems). Appropriate positive and negative controls were used throughout.
Hammer P.M., Kunder C.A., Howitt B.E, & Charville G.W. (2022). Well-differentiated lipomatous neoplasms with p53 alterations: a clinicopathological and molecular study of eight cases with features of atypical pleomorphic lipomatous tumour. Histopathology, 80(4), 656-664.
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3

Quantifying EZH2 and PTEN in Breast Cancer

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IHC was performed on formalin-fixed paraffin-embedded blanks from breast cancer cases from MSSM. Staining for EZH2 was performed and validated at the Molecular Cytology Core at MSKCC. Staining for PTEN was performed at Mount Sinai using the Leica-BOND automated IHC stainer. H&E slides for each case were provided with the blanks by MSSM. QurPath software was used for the analysis of IHC stain intensity quantification for PTEN and EZH260 (link).
Pappas K., Martin T.C., Wolfe A.L., Nguyen C.B., Su T., Jin J., Hibshoosh H, & Parsons R. (2021). NOTCH and EZH2 collaborate to repress PTEN expression in breast cancer. Communications Biology, 4, 312.
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4

Histological Analysis of Pancreatic Islets

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Muscle sections were cut at 4 µm for histological analysis in hematoxylin and eosin (H&E) staining. Islets were identified using an anti-islet 1 antibody (clone EPR10362, ab178400, 1:250 dilution, Abcam) with human pancreas used as a positive control. Blocking was done with 1× Animal-Free Blocker (SP-5030-250, Thermo Fisher Scientific) in 2% NGS (5425S, Cell Signaling Technology) diluted in TBST (J77500.K2, Thermo Fisher Scientific). The primary antibody was diluted in blocking solution at a titer of 1:1,000. Heat-mediated antigen retrieval was done on the Leica BOND using EDTA-based pH 9 solution (AR9640, Leica). A Leica BOND detection kit (DS9800) was used (post-primary step was omitted) for DAB chromogenic staining.
Hu X., White K., Olroyd A.G., DeJesus R., Dominguez A.A., Dowdle W.E., Friera A.M., Young C., Wells F., Chu E.Y., Ito C.E., Krishnapura H., Jain S., Ankala R., McGill T.J., Lin A., Egenberger K., Gagnon A., Michael Rukstalis J., Hogrebe N.J., Gattis C., Basco R., Millman J.R., Kievit P., Davis M.M., Lanier L.L., Connolly A.J., Deuse T, & Schrepfer S. (2023). Hypoimmune induced pluripotent stem cells survive long term in fully immunocompetent, allogeneic rhesus macaques. Nature Biotechnology, 42(3), 413-423.
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5

Immunohistochemical Staining for IgG4

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Immunohistochemical staining for IgG4 was performed on all cases. In addition, 10 histologically unremarkable oesophageal biopsies were examined to evaluate the presence and extent of background staining. All staining was performed on the Leica Bond automated platform (Cell Marque; Rocklin, CA, USA clone MRQ-44; dilution 1:25). A control sample, i.e. a lymph node with elevated numbers of IgG4-positive plasma cells, was included on each slide as an internal control. The biopsies were examined for immunoglobulin precipitate and the presence of IgG4-positive plasma cells.
Both the immunohistochemical slides and the H&E-stained slides were reviewed in a blinded manner.
Zukerberg L., Mahadevan K., Selig M, & Deshpande V. (2016). Oesophageal intrasquamous IgG4 deposits: an adjunctive marker to distinguish eosinophilic oesophagitis from reflux oesophagitis. Histopathology, 68(7), 968-976.
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6

Quantifying CD8+ and CD3+ T-Cell Density

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To determine CD8 þ and CD3 þ T-cell density, immunohistochemical studies were performed on a Leica Bond autostainer for CD3 (Dako, Carpinteria, CA; A0452; 1:100) or CD8 (Life Sciences Technologies, Waltham, MA; MS457s; 1:25) and 3,3'-diaminobenzidine chromogen and quantified using Aperio Scanscope AT Turbo, as described (Feldmeyer et al., 2016) (See Supplementary Figure S2 for a schematic as to this methodology).
Farah M., Reuben A., Spassova I., Yang R.K., Kubat L., Nagarajan P., Ning J., Li W., Aung P.P., Curry J.L., Torres-Cabala C.A., Hudgens C.W., Ugurel S., Schadendorf D., Gumbs C., Little L.D., Futreal A., Wistuba I.I., Prieto V.G., Wang L., Wong M.K., Wargo J.A., Becker J.C, & Tetzlaff M.T. (2020). T-Cell Repertoire in Combination with T-Cell Density Predicts Clinical Outcomes in Patients with Merkel Cell Carcinoma. The Journal of investigative dermatology, 140(11).
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