Context:

Thanks to the FEDER-FSE 2014-2020 Languedoc Roussillon Operational Program and co-funding from IBiSA, RHEM acquired in March 2018 an automated immunostaining and in situ hybridization system: the Ventana Roche Discovery Ultra.

Equipment:

This new system enables immunohistochemical (IHC) and immunofluorescent (IF) staining in single or multiplex formats. The advantages of this system are:

• 30 independent drawers, allowing up to 30 different protocols in a single cycle, with one protocol per slide.
• Even distribution of reagents across the slide, ensuring better staining uniformity thanks to regularly pulsed air flows.
• The possibility to have multiple tissue sections from different blocks on a single slide.
• Automated or manual deposition of antibodies and probes, reducing reagent dead volumes during optimization.
• Deposition of a small volume of antibodies and probes (100 µL/slide), lowering reaction costs.
• Control of antibody incubation temperature at 37°C.
• Full accessibility of the system, allowing the use of kits from any brand.
• Traceability ensuring all conditions applied to your slides are recorded.

With this equipment, we have access to the latest technological developments in IHC and fluorescent IHC, enabling multiplex staining in visible and fluorescence even with primary antibodies from the same species.

Applications:

The services offered with this system are as follows:

1) Execution of pre-validated stains on the system:

• Inflammatory cell markers: CD3e, PAX5, CD45, F4/80
• Proliferation or apoptosis markers: Ki67 (human or mouse), activated Casp3, phospho-H3
• Blood and lymphatic vessel markers: CD31, LYVE-1
• DNA damage markers: phospho-γH2AX
• Reporter tags or genes: GFP
• Tumor suppressors: P53
• Human cell markers: ALU probe, surface of intact mitochondria
• Signaling pathway markers: HES1
• Cell type markers: vimentin, Pan-CK, alpha-SMA

2) Development and automation of new single or multiplex immunostaining protocols upon request by a researcher or the platform steering committee.

To reduce costs, it is possible to place 2 sections from 2 different blocks on the same slide. Please contact us for guidance.

If you are interested in other markers, feel free to submit your development ideas.

New immunostaining protocols will be shared on the RHEM website or through the Biocampus newsletter.