This video analyzes the immune synapse formation between T cells in unpurified pan-leukocyte preparations and Staphylococcus aureus enterotoxin B, SEB-loaded Raji cells. CD3 and F-actin enrichment is observed in the immune synapse using imaging flow cytometry.
Protocol
All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines. 1. Preparation of Pan-leukocytes Draw 1 mL of peripheral blood from a healthy donor (or patient) in a heparinized syringe. Make sure to have approval by the responsible ethics committee for the blood donation. Mix 1 mL of human peripheral blood with 30 mL of ACK lysis buffer (150 mM NH4Cl, 1 mM KHCO3,…
Analysis of Immune Synapses between Human T Cells and SEB-Loaded Raji Cells using Imaging Flow Cytometry. J. Vis. Exp. (Pending Publication), e21665, doi: (2023).