Simple methods for quantifying super-resolved cortical actin

Evelyn Garlick, Emma L Faulkner, Stephen J Briddon, Steven G Thomas

Research output: Contribution to journalArticlepeer-review

Abstract

Cortical actin plays a key role in cell movement and division, but has also been implicated in the organisation of cell surface receptors such as G protein-coupled receptors. The actin mesh proximal to the inner membrane forms small fenced regions, or 'corrals', in which receptors can be constrained. Quantification of the actin mesh at the nanoscale has largely been attempted in single molecule datasets and electron micrographs. This work describes the development and validation of workflows for analysis of super resolved fixed cortical actin images obtained by Super Resolved Radial Fluctuations (SRRF), Structured Illumination Microscopy (3D-SIM) and Expansion Microscopy (ExM). SRRF analysis was used to show a significant increase in corral area when treating cells with the actin disrupting agent cytochalasin D (increase of 0.31 µm2 ± 0.04 SEM), and ExM analysis allowed for the quantitation of actin filament densities. Thus, this work allows complex actin networks to be quantified from super-resolved images and is amenable to both fixed and live cell imaging.

Original languageEnglish
Article number2715
JournalScientific Reports
Volume12
Issue number1
DOIs
Publication statusPublished - 17 Feb 2022

Bibliographical note

© 2022. The Author(s).

Keywords

  • A549 Cells
  • Actin Cytoskeleton/chemistry
  • Actins/analysis
  • Cell Membrane/chemistry
  • Cytochalasin D/pharmacology
  • Humans
  • Image Processing, Computer-Assisted/methods
  • Microscopy, Fluorescence/methods

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