Analyzing trogocytosis of T lymphocytes by flow cytometry and confocal microscopy

Abstract

Summary: Here, we present a protocol to examine the mechanisms underlying the intercellular transfer of transmembrane molecules, termed trogocytosis, and the fate of transferred molecules. We describe the steps needed from T lymphocyte isolation, via co-culture with cells expressing the ligand of interest, to cell harvest and subsequent staining for flow cytometry and confocal microscopy. Furthermore, we showcase critical parameters and pitfalls, which allow easy adaptation of the protocol to investigate trogocytosis of various cell surface receptors in different cell types.For complete details on the use and execution of this protocol, please refer to Zink and Rohr.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Cell Biology
• Flow Cytometry/Mass Cytometry
• Cell Membrane
• Cell-based Assays
• Immunology