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Immunology and Infection

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Nanyang Technological University

Cortical Actin Flow in T Cells Quantified by Spatio-temporal Image Correlation Spectroscopy of Structured Illumination Microscopy Data

9.7K Views

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09:09 min

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December 17th, 2015

DOI :

10.3791/53749-v

December 17th, 2015

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George Ashdown¹, Elvis Pandžić³, Andrew Cope², Paul Wiseman⁴, Dylan Owen¹

¹Department of Physics and Randall Division of Cell and Molecular Biophysics, King's College London, ²Academic Department of Rheumatology, Centre for Molecular and Cellular Biology of Inflammation, Division of Immunology, Infection and Inflammatory Disease, King's College London, ³ARC Centre for Advanced Molecular Imaging, Australian Centre for NanoMedicine, University of New South Wales Australia, ⁴Departments of Chemistry and Physic, McGill University

Summary

To investigate flow velocities and directionality of filamentous-actin at the T cell immunological synapse, live-cell super-resolution imaging is combined with total internal reflection fluorescence and quantified with spatio-temporal image correlation spectroscopy.

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Keywords Actin Flow

Filamentous Actin

Immunological Synapse

Structured Illumination Microscopy

Spatio temporal Image Correlation Spectroscopy

Super resolution Imaging

Retrograde Flow

Centripetal Flow

Sub diffraction

Molecular Distributions

Vesicle Dynamics

Protein Clustering

Chapters in this video

0:05

Title

0:50

Microscope Set-up

3:20

Cell Imaging

4:44

Structured Illumination Microscopy (SIM) Image Reconstruction and Spatio-temporal Image Correlation Spectroscopy (STICS) Analysis

7:12

Results: Representative Structured Illumination Data and STICS Output

8:36

Conclusion

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