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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
PubMed
Summary
This summary is machine-generated.

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.

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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.