Live‐cell super‐resolution imaging of actin using <scp>LifeAct</scp> ‐14 with a <scp>PAINT</scp> ‐based approach
Haresh Bhaskar, Dirk‐Jan Kleinjan, Curran Oi, Zoe Gidden, Susan J. Rosser, Mathew H. Horrocks, Lynne Regan
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.
Topics & Concepts
ActinLive cell imagingSuperresolutionEukaryotic cellCell biologyCellChemistryBiophysicsBiologyComputer scienceBiochemistryArtificial intelligenceImage (mathematics)Advanced Fluorescence Microscopy TechniquesCell Image Analysis TechniquesAdvanced Electron Microscopy Techniques and Applications