Molecular Flow Quantified beyond the Diffraction Limit by Spatiotemporal Image Correlation of Structured Illumination Microscopy Data

We combine total internal reflection fluorescence structured illumination microscopy with spatiotemporal image correlation spectroscopy to quantify the flow velocities and directionality of filamentous-actin at the T cell immunological synapse. These techniques demonstrate it is possible to image re...

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Published inBiophysical journal Vol. 107; no. 9; pp. L21 - L23
Main Authors Ashdown, George W., Cope, Andrew, Wiseman, Paul W., Owen, Dylan M.
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 04.11.2014
Biophysical Society
The Biophysical Society
Subjects
Actins - metabolism
Biophysical Letter
Biophysics
CD4 Antigens - metabolism
Flow velocity
Fluorescence
Humans
Image Processing, Computer-Assisted - methods
Immunology
Jurkat Cells
Microscopy
Microscopy, Fluorescence - methods
Molecular Imaging - methods
Spectrum analysis
Spectrum Analysis - methods
T cell receptors
T-Lymphocytes - metabolism
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Summary:We combine total internal reflection fluorescence structured illumination microscopy with spatiotemporal image correlation spectroscopy to quantify the flow velocities and directionality of filamentous-actin at the T cell immunological synapse. These techniques demonstrate it is possible to image retrograde flow of filamentous-actin at superresolution and provide flow quantification in the form of velocity histograms and flow vector maps. The flow was found to be retrograde and radially directed throughout the periphery of T-cells during synapse formation.
Bibliography:ObjectType-Article-1
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content type line 23
ISSN:0006-3495
1542-0086
DOI:10.1016/j.bpj.2014.09.018