期刊
BIOPHYSICAL JOURNAL
卷 95, 期 10, 页码 L66-L68出版社
BIOPHYSICAL SOC
DOI: 10.1529/biophysj.108.143198
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资金
- Biotechnology and Biological Sciences Research Council
- Department of Trade and Industry
- Lister Institute Research Prize
- Medical Research Council
- Royal Society
- Engineering and Physical Sciences Research Council
- MRC [G0500563] Funding Source: UKRI
- Medical Research Council [G0500563] Funding Source: researchfish
Imaging in any plane other than horizontal in a microscope typically requires a reconstruction from multiple optical slices that significantly decreases the spatial and temporal resolution that can be achieved. This can limit the precision with which molecular events can be detected, for example, at intercellular contacts. This has been a major issue for the imaging of immune synapses between live cells, which has generally required the reconstruction of en face intercellular synapses, yielding spatial resolution significantly above the diffraction limit and updating at only a few frames per minute. Strategies to address this issue have usually involved using artificial activating substrates such as antibody-coated slides or supported planar lipid bilayers, but synapses with these surrogate stimuli may not wholly resemble immune synapses between two cells. Here, we combine optical tweezers and confocal microscopy to realize generally applicable, high-speed, high-resolution imaging of almost any arbitrary plane of interest. Applied to imaging immune synapses in live-cell conjugates, this has enabled the characterization of complex behavior of highly dynamic clusters of T cell receptors at the T cell/antigen-presenting cell intercellular immune synapse and revealed the presence of numerous, highly dynamic long receptor-rich filopodial structures within inhibitory Natural Killer cell immune synapses.
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