Protein ScienceSynthetic BiologyPeer-reviewed journal article
Live-cell super-resolution imaging of actin using LifeAct-14 with a PAINT-based approach
Haresh BhaskariD, Dirk‐Jan Kleinjan, Curran OiiD, Zoe GiddeniD, Susan J. RosseriD, Mathew H. HorrocksiD, Lynne ReganiD
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.
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