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Biophys J. 2014 Nov 4;107(9):L21-3. doi: 10.1016/j.bpj.2014.09.018.

Molecular flow quantified beyond the diffraction limit by spatiotemporal image correlation of structured illumination microscopy data.

Author information

1
Department of Physics and Randall Division of Cell and Molecular Biophysics, King's College London, London, United Kingdom.
2
Academic Department of Rheumatology, Centre for Molecular and Cellular Biology of Inflammation, Division of Immunology, Infection and Inflammatory Disease, King's College London, London, United Kingdom.
3
Departments of Chemistry and Physics, McGill University, Montreal, Quebec, Canada.
4
Department of Physics and Randall Division of Cell and Molecular Biophysics, King's College London, London, United Kingdom. Electronic address: dylan.owen@kcl.ac.uk.

Abstract

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.

PMID:
25418107
PMCID:
PMC4223199
DOI:
10.1016/j.bpj.2014.09.018
[Indexed for MEDLINE]
Free PMC Article

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