Abstract:
: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.
journal_name
Biophys Jjournal_title
Biophysical journalauthors
Oddos S,Dunsby C,Purbhoo MA,Chauveau A,Owen DM,Neil MA,Davis DM,French PMdoi
10.1529/biophysj.108.143198subject
Has Abstractpub_date
2008-11-15 00:00:00pages
L66-8issue
10eissn
0006-3495issn
1542-0086pii
S0006-3495(08)78589-8journal_volume
95pub_type
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