Live‐cell super‐resolution imaging of actin using LifeAct‐14 with a PAINT‐based approach |
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| Authors: | Haresh Bhaskar,Dirk‐ Jan Kleinjan,Curran Oi,Zoe Gidden,Susan J. Rosser,Mathew H. Horrocks,Lynne Regan |
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| Affiliation: | 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 |
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| 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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| Keywords: | actin, LifeAct, live‐ cell imaging, single‐ molecule localization microscopy (SMLM), super‐ resolution (SR) imaging |
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