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Updated: Oct 3, 2025
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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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Simple methods for quantifying super-resolved cortical actin
Evelyn Garlick1,2, Emma L Faulkner1,2, Stephen J Briddon2,3
1Institute of Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK.
Scientific Reports
|February 18, 2022
Summary
This study quantifies nanoscale cortical actin networks using super-resolution microscopy. New workflows enable precise measurement of actin corrals and filament densities, advancing cell biology research.
Area of Science:
- Cell Biology
- Biophysics
- Microscopy
Background:
- Cortical actin structures, including actin corrals, regulate cell surface receptor organization and mobility.
- Existing nanoscale quantification methods for actin meshes are limited.
Purpose of the Study:
- To develop and validate workflows for analyzing super-resolved images of cortical actin.
- To enable quantitative analysis of nanoscale actin networks in fixed and live cells.
Main Methods:
- Super-Resolved Radial Fluctuations (SRRF) microscopy
- 3D Structured Illumination Microscopy (3D-SIM)
- Expansion Microscopy (ExM)
- Analysis of super-resolved fixed cortical actin images
Main Results:
- Developed and validated workflows for SRRF, 3D-SIM, and ExM image analysis.
- Demonstrated a significant increase in corral area upon treatment with cytochalasin D (0.31 µm² ± 0.04 SEM) using SRRF.
- Quantified actin filament densities using ExM.
Conclusions:
- Established robust methods for quantifying complex nanoscale actin networks from super-resolved images.
- The developed workflows are applicable to both fixed and live cell imaging.
- Provides new tools for understanding the role of cortical actin in cellular processes.
