Bond max platform

Manufactured by Leica

The Bond Max platform is a state-of-the-art automated slide staining system designed for immunohistochemistry and in situ hybridization applications. It offers a high level of automation and standardization to streamline laboratory workflows.

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Lab products found in correlation

Axioskop microscope, by Zeiss (1 mentions) Ab28364, by Abcam (1 mentions) Ab92696, by Abcam (1 mentions) Anti nalp3 cias1 nlrp3, by Abcam (1 mentions)

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Bond Max (596 citations) Bond platform (16 citations)

3 protocols using bond max platform

Co-expression Analysis of miR-155 Targets

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All sections were dealt with blindly using an automated Leica Bond Max platform. The coverslips of the miR-155 stained slides were removed and the tissues tested with antibodies of interest for co-expression analyses. The antibodies used for immunohistochemistry were: MFSD2A (#PA5–21049, Invitrogen); ChAT (#AB144P, Millipore); Neuron-Specific Enolase (BML-NA1501–0100, Enzo Life Sciences); CD31 (ab28364, Abcam), and pyruvate dehydrogenase (ab92696, ABCAM).
Co-expression analyses were done using the Nuance system (CRI) as previously published [16 (link)]. In brief, a given tissue was tested for two different targets using fast red, NBT/BCIP or DAB as the chromogens. The results were then analyzed by the Nuance and InForm systems with a with the Zeiss Axioskop microscope to determine what percentage of cells were expressing the two targets of interest.

Awad H., Bratasz A., Nuovo G., Burry R., Meng X., Kelani H., Brown M., Ramadan M.E., Williams J., Bouhliqah L., Popovich P.G., Guan Z., Mcallister C., Corcoran S.E., Kaspar B., Basso D.M., Otero J.J., Kirsch C., Davis I.C., Croce C.M., Michaille J.J, & Tili E. (2018). MiR-155 deletion reduces ischemia-induced paralysis in an aortic aneurysm repair mouse model: Utility of immunohistochemistry and histopathology in understanding etiology of spinal cord paralysis. Annals of diagnostic pathology, 36, 12-20.

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Optimized Immunostaining of SARS-CoV-2 Proteins

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Our immunohistochemistry method for the detection of SARS-CoV-2 proteins has been published [2 (link)]. The automated Leica Bond Max platform was used with DAB as the chromogen. The optimal conditions included antigen retrieval for 30 min with the EDTA solution from Leica, dilutions of 1:4000 (spike Ab), 1:500 (membrane Ab), 1:250 (envelope Ab, each from ProSci, Poway, CA) and the use of the horseradish peroxidase conjugate from Enzo Life Sciences in place of the equivalent product from Leica as this reduced background [11 (link),12 ]. In selected cases the chromogen Fast Red (with the alkaline phosphatase reporter enzyme) was used in place of DAB by using the Leica Fast Red kit. Positive and negative controls were lung tissues from people who had died of COVID-19 and normal lung tissue obtained prior to 2018.

Nuovo G., Tili E., Suster D., Matys E., Hupp L, & Magro C. (2020). Strong homology between SARS-CoV-2 envelope protein and a Mycobacterium sp. antigen allows rapid diagnosis of Mycobacterial infections and may provide specific anti-SARS-CoV-2 immunity via the BCG vaccine. Annals of Diagnostic Pathology, 48, 151600.

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NLRP3 and Caspase-1 Immunohistochemistry

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Immunostains were performed on 5 μM formalin-fixed paraffin-embedded histologic sections using the automated Leica Bond-Max platform. Anti-NALP3/CIAS1 (NLRP3) (Abcam, Cambridge, MA) primary antibodies were detected using a post-primary biotin-free alkaline phosphate polymer and fast red chromogen (Leica Biosystems, Buffalo Grove, IL). Rabbit monoclonal anti-CASP1 antibodies were used for IHC (Cell Signaling Technology, Danvers, MA) and polyclonal rabbit anti-CASP1 anti-sera utilized for SDS-PAGE was generously provided by Dr. Gabriel Nunez (University of Michigan Medical School, Ann Arbor, MI).

von Herrmann K.M., Salas L.A., Martinez E.M., Young A.L., Howard J.M., Feldman M.S., Christensen B.C., Wilkins O.M., Lee S.L., Hickey W.F, & Havrda M.C. (2018). NLRP3 expression in mesencephalic neurons and characterization of a rare NLRP3 polymorphism associated with decreased risk of Parkinson’s disease. NPJ Parkinson's Disease, 4, 24.

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