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

Requirements:

  • 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

Application form and further details

CALL CLOSED

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