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09:09
Cortical Actin Flow in T Cells Quantified by Spatio-temporal Image Correlation Spectroscopy of Structured Illumination Microscopy Data
Published on: December 17, 2015
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Molecular flow quantified beyond the diffraction limit by spatiotemporal image correlation of structured illumination
George W Ashdown1, Andrew Cope2, Paul W Wiseman3
1Department of Physics and Randall Division of Cell and Molecular Biophysics, King's College London, London, United Kingdom.
Biophysical Journal
|November 25, 2014
Summary
Researchers quantified filamentous-actin flow at T-cell synapses using advanced microscopy and spectroscopy. Actin filaments move retrograde and radially in T-cells during synapse formation, providing insights into immune cell function.
Area of Science:
- Cell Biology
- Immunology
- Biophysics
Background:
- The T-cell immunological synapse is crucial for adaptive immunity.
- Understanding the dynamics of the actin cytoskeleton is key to T-cell function.
Purpose of the Study:
- To quantify the flow velocities and directionality of filamentous-actin at the T-cell immunological synapse.
- To visualize actin dynamics at super-resolution during immune cell interactions.
Main Methods:
- Combined total internal reflection fluorescence structured illumination microscopy (TIRF-SIM) with spatiotemporal image correlation spectroscopy (4D-SPIM).
- Utilized super-resolution microscopy to image actin.
- Employed image correlation spectroscopy for flow quantification.
Main Results:
- Achieved super-resolution imaging of filamentous-actin retrograde flow.
- Provided quantitative flow data via velocity histograms and flow vector maps.
- Observed retrograde and radially directed flow of actin throughout the T-cell periphery during synapse formation.
Conclusions:
- The combined techniques enable super-resolution imaging and quantification of actin dynamics.
- Actin flow is retrograde and radially directed at the T-cell synapse periphery.
- These findings offer insights into the mechanics of T-cell activation and immune response.
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