Live-cell super-resolution imaging of actin using LifeAct-14 with a PAINT-based approach

Haresh Bhaskar; Dirk-Jan Kleinjan; Curran Oi; Zoe Gidden; Susan J Rosser; Mathew H Horrocks; Lynne Regan

doi:10.1002/pro.4558

Live-cell super-resolution imaging of actin using LifeAct-14 with a PAINT-based approach

Protein Sci. 2023 Feb;32(2):e4558. doi: 10.1002/pro.4558.

Authors

Haresh Bhaskar^{1

2}, Dirk-Jan Kleinjan³, Curran Oi⁴, Zoe Gidden^{1

2}, Susan J Rosser³, Mathew H Horrocks², Lynne Regan¹

Affiliations

¹ The School of Biological Sciences, University of Edinburgh, Edinburgh, UK.
² EaStCHEM School of Chemistry, University of Edinburgh, Edinburgh, UK.
³ Centre for Synthetic and Systems Biology and UK Centre for Mammalian Synthetic Biology, School of Biological Sciences, University of Edinburgh, Edinburgh, UK.
⁴ Department of Genome Sciences, University of Washington, Seattle, Washington, USA.

Abstract

We present direct-LIVE-PAINT, an easy-to-implement approach for the nanoscopic imaging of protein structures in live cells using labeled binding peptides. We demonstrate the feasibility of direct-LIVE-PAINT with an actin-binding peptide fused to EGFP, the location of which can be accurately determined as it transiently binds to actin filaments. We show that direct-LIVE-PAINT can be used to image actin structures below the diffraction-limit of light and have used it to observe the dynamic nature of actin in live cells. We envisage a similar approach could be applied to imaging other proteins within live mammalian cells.

Keywords: LifeAct; actin; live-cell imaging; single-molecule localization microscopy (SMLM); super-resolution (SR) imaging.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Actin Cytoskeleton*
Actins* / metabolism
Animals
Mammals
Protein Binding

Substances

Actins

Abstract

Publication types

MeSH terms

Substances

Grants and funding