Live Cell Linear Dichroism Imaging Reveals Extensive Membrane Ruffling within the Docking Structure of Natural Killer Cell Immune Synapses |
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Authors: | Richard KP Benninger Stephen Young Fiona J Culley Mark AA Neil Paul MW French Björn Önfelt |
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Institution: | † Department of Physics, Imperial College London, London, United Kingdom ‡ Cell Physics, Department of Applied Physics, Royal Institute of Technology, Stockholm, Sweden § Division of Cell and Molecular Biology, Imperial College London, London, United Kingdom ¶ Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark ‖ Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden |
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Abstract: | We have applied fluorescence imaging of two-photon linear dichroism to measure the subresolution organization of the cell membrane during formation of the activating (cytolytic) natural killer (NK) cell immune synapse (IS). This approach revealed that the NK cell plasma membrane is convoluted into ruffles at the periphery, but not in the center of a mature cytolytic NK cell IS. Time-lapse imaging showed that the membrane ruffles formed at the initial point of contact between NK cells and target cells and then spread radialy across the intercellular contact as the size of the IS increased, becoming absent from the center of the mature synapse. Understanding the role of such extensive membrane ruffling in the assembly of cytolytic synapses is an intriguing new goal. |
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