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Bond max instrument

Manufactured by Leica
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The Bond-Max instrument is a piece of laboratory equipment designed for automated nucleic acid hybridization. It performs procedures such as in situ hybridization and immunohistochemistry for research and diagnostic applications.

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Lab products found in correlation

Bond polymer refine detection kit, by Leica (3 mentions) Bond polymer refine detection reagents, by Leica (2 mentions) Human vimentin, by Cell Signaling Technology (2 mentions) Optiview dab ihc detection kit, by Roche (1 mentions) Benchmark gx, by Roche (1 mentions) Cyclin d1 clone sp4, by Thermo Fisher Scientific (1 mentions) P53 clone do 7, by Agilent Technologies (1 mentions) Bond polymer refine detection system, by Leica (1 mentions) Glypican 3, by Biocare Medical (1 mentions) P s6k, by Cell Signaling Technology (1 mentions)

Close spelling variants of this product

Bond Max (596 citations) Bond instrument (2 citations)

8 protocols using bond max instrument

1

Postmortem Biopsy Procedure for COVID-19

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With permission from the patients' families, postmortem needle core biopsies were performed on visceral organs including the lungs, liver, and heart within an hour after death in a negative air isolation ward. The procedures were performed without ultrasound guidance, but the patients' last radiographic images and surface anatomic landmarks were used as references. The tissues were received fixed in neutral buffered formalin for over 24 h, and then routinely processed under standard biosafety measures. Hematoxylin and eosin-stained sections were prepared and slides were examined by two pathologists (SFT and SYX). Immunohistochemistry (IHC) staining was performed in a liver specimen from Case 1, who had a history of CLL. It was used to verify subsets of the small lymphocytes found in portal tracts, using antibodies against CD20, CD3, CD5, CD23, CD4, and CD8 (Agilent Technologies, US). All antibodies were used in prediluted form, and IHC was performed using the automated Leica Bond-Max instrument.
Tian S., Xiong Y., Liu H., Niu L., Guo J., Liao M, & Xiao S.Y. (2020). Pathological study of the 2019 novel coronavirus disease (COVID-19) through postmortem core biopsies. Modern Pathology, 33(6), 1007-1014.
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2

Formalin-Fixed Lung Tissue Analysis

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Lungs were harvested from euthanized mice and fixed in 10% formalin overnight. The formalin-fixed samples were sent to the Cambridge University Hospital Human Research Tissue Bank for paraffin-embedding and sectioning. Human vimentin (Cell Signaling Technology; Cat. 5741, 1:100) staining was performed in a Bond-Max instrument (Leica) using Bond Polymer Refine Detection reagents (Leica) according to the manufacturer’s protocol (IHC Protocol F).
Syafruddin S.E., Rodrigues P., Vojtasova E., Patel S.A., Zaini M.N., Burge J., Warren A.Y., Stewart G.D., Eisen T., Bihary D., Samarajiwa S.A, & Vanharanta S. (2019). A KLF6-driven transcriptional network links lipid homeostasis and tumour growth in renal carcinoma. Nature Communications, 10, 1152.
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3

Immunohistochemistry Staining Protocols

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Immunohistochemistry staining was performed on Ventana BenchMark GX using OptiView DAB IHC Detection Kit (Ventana Medical Systems, Tucson USA) whereas EBER immunohistochemical staining was performed on Bond-Max instrument (Leica, Newcastle Upon Tyne, UK) using Bond Polymer Refine Detection kit (Leica Biosystems, Newcastle Upon, United Kingdom). The test protocols and the antibodies used for immunohistochemistry staining are shown in Table 1.
Ting C.Y., Chang K.M., Kuan J.W., Sathar J., Chew L.P., Wong O.L., Yusuf Y., Wong L., Samsudin A.T., Pana M.N., Lee S.K., Gopal N.S., Puri R., Ong T.C., Bahari S.K., Goh A.S, & Teoh C.S. (2019). Clinical Significance of BCL2, C-MYC, and BCL6 Genetic Abnormalities, Epstein-Barr Virus Infection, CD5 Protein Expression, Germinal Center B Cell/Non-Germinal Center B-Cell Subtypes, Co-expression of MYC/BCL2 Proteins and Co-expression of MYC/BCL2/BCL6 Proteins in Diffuse Large B-Cell Lymphoma: A Clinical and Pathological Correlation Study of 120 Patients. International Journal of Medical Sciences, 16(4), 556-566.
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4

Histological Analysis of Lungs and Tumors

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Lungs and subcutaneous tumors were collected and fixed overnight in 4% paraformaldehyde, washed, embedded in paraffin and sectioned. H&E staining was performed by standard methods. Human Vimentin (Cell Signaling, Cat. 5741, 1:100) staining was performed in a Bond-Max instrument (Leica) using Bond Polymer Refine Detection reagents (Leica) according to the manufacturer’s protocol (IHC Protocol F).
Rodrigues P., Patel S.A., Harewood L., Olan I., Vojtasova E., Syafruddin S.E., Zaini M.N., Richardson E.K., Burge J., Warren A.Y., Stewart G.D., Saeb-Parsy K., Samarajiwa S.A, & Vanharanta S. (2018). NF-kappaB-dependent lymphoid enhancer co-option promotes renal carcinoma metastasis. Cancer discovery, 8(7), 850-865.
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5

Tissue Microarray Immunohistochemical Profiling

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Three-μm thick tissue microarray sections were deparaffinized followed by antigen retrieval (EDTA pH 9.0, 95 °C, 30 min). Stainings were performed on a Leica BOND MAX instrument (Leica) using the Bond Polymer Refine detection kit (Leica) and the following primary antibodies: p16INK4a (clone E6H4, dilution 1:10, 30 min; MTM Laboratories, Heidelberg, Germany), pRB (clone 13A10, dilution 1:100, 15 min; Novocastra), cyclin D1 (clone SP4, dilution 1:50, 30 min; ThermoScientific), and p53 (clone DO-7, dilution 1:150, 30 min; Dako). Marker expression was scored as high or low based on the proportion of positive carcinoma cells (high p16INKa: more than 70% of tumor cells; high pRB, cyclin D1, or p53: more than 25% of tumor cell nuclei) [19 (link), 21 (link)].
Plath M., Broglie M.A., Förbs D., Stoeckli S.J, & Jochum W. (2018). Prognostic significance of cell cycle-associated proteins p16, pRB, cyclin D1 and p53 in resected oropharyngeal carcinoma. Journal of Otolaryngology - Head & Neck Surgery, 47, 53.
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6

Immunohistochemical Profiling of Tumor Markers

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One to two representative formalin-fixed, paraffin-embedded blocks from each case were selected for immunohistochemical staining. Automated immunohistochemical staining was performed with a Leica Bond Max Instrument (Leica, Buffalo Grove, IL, USA). Tissue sections (4 μm) were deparaffinized, rehydrated and treated with 3% hydrogen peroxide for 15 min to quench endogenous peroxidase. Antigen retrieval was performed using a bond epitope retrieval solution [an ethylenediamine tetraacetic acid (EDTA)-based buffer and surfactant] at pH 9.0 for 20 min. The slides were first incubated with one of the following primary antibodies: beta-catenin (3.5 mg/l, Leica), glypican-3 (1:50; Biocare, Concord, CA, USA), spalt-like transcription factor 4 (SALL4) (ready to use; clone 6E3; Cell Marque, Rocklin, CA, USA) and telomerase reverse transcriptase (TERT) (A-6) (1:50; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), following the manufacturers’ instructions. After incubation with either an antimouse or antirabbit secondary antibody, a Bond Polymer Refine Detection system (Leica) was used for single brown colour staining with 3,3′ diaminobenzidine (DAB) chromogen. Appropriate positive and negative controls were included and evaluated with the specimens tested.
Zhou S., Venkatramani R., Gupta S., Wang K., Stein J.E., Wang L, & Mascarenhas L. (2017). Hepatocellular malignant neoplasm, NOS: a clinicopathological study of 11 cases from a single institution. Histopathology, 71(5), 813-822.
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7

Immunohistochemical Analysis of Signaling Proteins

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Sections for IHC were stained with antibodies for the signaling proteins, IGF1Rb (Leica Biosystems, Buffalo Grove, IL) and p-S6K (Cell Signaling, Danvers, MA) according to the manufacturer’s specifications. Automated immunostaining was performed with a Leica Bond Max instrument (Leica, Richmond, Illinois). After deparaffinization and antigen retrieval, tumor microarray and control slides were sequentially incubated with the primary antibody, a secondary antibody, a polymer conjugate, and a coloring reagent that gives a brown color staining. Separate positive controls were made from breast carcinoma, all of which were previously shown to exhibit expression of the studied proteins. Negative controls were stained similarly except for omission of primary antibodies. Staining of vascular endothelial cells served as internal positive control.
Gonzalez E., Bui M, & Ahmed A.A. (2020). IGF1R immunohistochemistry in Ewing’s sarcoma as predictor of response to targeted therapy. International Journal of Health Sciences, 14(4), 17-21.
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8

Ki-67 Immunohistochemistry Protocol for Tumor Analysis

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Ki-67 assessments were performed by the same two laboratory technicians over the whole study period. It was performed in a manual way for all cases. Two-μm thick formalin-fixed, paraffin-embedded tissue sections were deparaffinized followed by antigen retrieval (EDTA pH 9.0, 95 °C, 30 min). Ki-67 stainings were performed on a Leica BOND MAX instrument (Leica) using mouse monoclonal Ki-67 antibody (clone MIB1, dilution 1:80, 30 min; Dako) and the Bond Polymer Refine detection kit (Leica). Ki-67 index was defined as the percentage of invasive carcinoma cells with nuclear Ki-67 staining. For each tumor, at least 1′000 carcinoma cells in hot spot areas were scored. In the literature two ways of counting Ki-67 index are discussed: average counting and condensed proliferative areas, so called ‘hot spot’- counting[20 (link),21 (link)]. As in Switzerland the guidelines recommend counting hot spots we decided to follow this recommendation as it is traditionally done in our institution [21 (link),22 ].
Maranta A.F., Broder S., Fritzsche C., Knauer M., Thürlimann B., Jochum W, & Ruhstaller T. (2020). Do YOU know the Ki-67 index of your breast cancer patients? Knowledge of your institution’s Ki-67 index distribution and its robustness is essential for decision-making in early breast cancer. The Breast : official journal of the European Society of Mastology, 51, 120-126.
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