STSM: Immunostaining of circulating and tissue-infiltrating neutrophils to facilitate neutrophil phenotypic and functional biomarker development in DACI
STSM CALL - Standardization of staining protocols for tissue neutrophils - NOW CLOSED
Host Institution: UCLouvain, Brussels, Belgium
Supervisor/Coordinator: Caroline Bouzin
Instructors: Caroline Bouzin, Aurélie Daumerie, Pierre Michel
Timeframe: 19th June to 23rd June 2023
Attendees: 6x WG 2 members
Purpose: Based on our expertise in multiplex immunofluorescence staining of FFPE sections, we will fine tune a 6-plex panel dedicated to neutrophil subsets detection. Each primary antibody will first be tested in standard IHC-DAB to confirm previously established concentrations, on FFPE sections of lung and head and neck cancer. Each antibody will then be tested in fluorescence at each position (1 to 6) of the multiplex using the tyramide signal amplification (TSA) method. This experiment aims at finding the best combination, some antigens being destroyed after several cycles while others are better exposed. Slides will then be digitalized using a slide scanner.
Tasks: Our aim is to optimize a combination of neutrophil markers suitable for FFPE sections (specific staining, good signal-to-noise ratio, no spill-over).
Selection Criteria: WG 2 members: Expertise in myeloid cell immunology with an interest in tissue sections immunostaining
- Availability from June 19 at 10:00 until June 23 at 13:00.
- Notebook / laptop
- Primary antibodies to be included in the multiplex
Expected Outcome: Based on the optimization performed during the STSM, we will run the chosen combination of antibodies on FFPE sections from different DACI samples to hopefully propose an optimal workflow.
Application Deadline: 1st May 2023