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M. Fölser, V. Motsch, R. Platzer, J. Huppa, G. Schütz:
"A multimodal platform for simultaneous T cell imaging, defined activation, and mechanobiological characterization";
Cells, 10 (2021), 23501 - 23512.

T-cell antigen recognition is accompanied by extensive morphological rearrangements ofthe contact zone between the T-cell and the antigen-presenting cell (APC). This process involvesbinding of the T-cell receptor (TCR) complex to antigenic peptides presented via MHC on the APCsurface, the interaction of costimulatory and adhesion proteins, remodeling of the actin cytoskeleton,and the initiation of downstream signaling processes such as the release of intracellular calcium.However, multiparametric time-resolved analysis of these processes is hampered by the difficultyin recording the different readout modalities at high quality in parallel. In this study, we present aplatform for simultaneous quantification of TCR distribution via total internal reflection fluorescencemicroscopy, of intracellular calcium levels, and of T-cell-exerted forces via atomic force microscopy(AFM). In our method, AFM cantilevers were used to bring single T-cells into contact with theactivating surface. We designed the platform specifically to enable the study of T-cell triggering viafunctionalized fluid-supported lipid bilayers, which represent a widely accepted model system tostimulate T-cells in an antigen-specific manner. In this paper, we showcase the possibilities of thisplatform using primary transgenic T-cells triggered specifically via their cognate antigen presentedby MHCII.

T-cell; immunological synapse; atomic force microscopy; total internal fluorescencemicroscopy; calcium imaging

http://dx.doi.org/10.3390/cells10020235