Comparing lifeact and phalloidin for super-resolution imaging of actin in fixed cells

PLoS One. 2021 Jan 28;16(1):e0246138. doi: 10.1371/journal.pone.0246138. eCollection 2021.

Abstract

Visualizing actin filaments in fixed cells is of great interest for a variety of topics in cell biology such as cell division, cell movement, and cell signaling. We investigated the possibility of replacing phalloidin, the standard reagent for super-resolution imaging of F-actin in fixed cells, with the actin binding peptide 'lifeact'. We compared the labels for use in single molecule based super-resolution microscopy, where AlexaFluor 647 labeled phalloidin was used in a dSTORM modality and Atto 655 labeled lifeact was used in a single molecule imaging, reversible binding modality. We found that imaging with lifeact had a comparable resolution in reconstructed images and provided several advantages over phalloidin including lower costs, the ability to image multiple regions of interest on a coverslip without degradation, simplified sequential super-resolution imaging, and more continuous labeling of thin filaments.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actin Cytoskeleton / chemistry
  • Actin Cytoskeleton / pathology*
  • Carbocyanines / chemistry*
  • HeLa Cells
  • Humans
  • Microscopy, Fluorescence
  • Phalloidine / chemistry*

Substances

  • Alexa Fluor 647
  • Carbocyanines
  • Phalloidine

Associated data

  • Dryad/10.5061/dryad.xsj3tx9cn