STSM: Establishing marker panels for myeloid cells in chronic inflammatory diseases
6-plex image: © Espen Sønderaal Bækkevold/Mye-InfoBank 2023
Host Institution: Oslo University Hospital, Norway
Supervisor/Coordinator: Espen S. Bækkevold
Instructors: Espen S. Bækkevold, Hogne Rød Nilsen, Kjersti Hagen
Timeframe: 20th September to 22nd September 2023
Attendees: On-site: 8-10x WG 2 & 3 members/instructor(s)
Purpose: One main output of WG 3 MYE-DISEASE is to facilitate the transformation of complex molecular data on myeloid cell signatures into biomarker panels employable in clinical diagnostic applications. To this end, immunostainings of tissue antigens offers the possibility to employ such panels in large collections of biosamples across research labs, and are readily compatible with routine pathological examinations.
Based on the initial testing of selected antibodies in the “cell-type-centric” STSMs, we will compile and test 6-plex marker panels on 4-5 tissue types (chronic inflammation and cancer) using immunofluorescence on FFPE-sections.
Tasks: We plan to start from fresh surgical resections (depending on availability) and cover all steps from dissection, fixation, paraffin embedding and sectioning. Sections will then be stained with an automated immunohistochemistry platform (Ventana Discovery) using Opal Fluorophore Reagent Packs (Akoya) for detection, and finally imaged on a PhenoImager (Akoya). Scanned images will be distributed within the Action for processing by interested members.
The workshop will start in the morning (~10 am) Wednesday September 20th with tissue preparation and sectioning (and a dinner + drinks), Thursday 21st (until quite late) will be focusing on the staining procedure, and Friday will finish in the afternoon (~3 pm) with presentations from attendees and a wrap-up.
Selection Criteria: Preferably some prior experience in immunohistochemistry or similar techniques, and some capacities (or ambitions) to implement multiplex immunofluorescence in their home labs.
Expected Outcome: Based on our work during the STSM, we will establish and test protocols for 6-plex immunofluorescence stainings (with 3-4 cell type-specific “backbone markers” and 2-3 “functional markers”) which can be implemented in the Action members labs for scaled analyses of tissue specimens from relevant disease specimens.
Application Deadline: 15th August 2023