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

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.

SUBMITTER: Bhaskar H

PROVIDER: S-EPMC9878614 | biostudies-literature | 2023 Feb

REPOSITORIES: biostudies-literature

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

Bhaskar Haresh H Kleinjan Dirk-Jan DJ Oi Curran C Gidden Zoe Z Rosser Susan J SJ Horrocks Mathew H MH Regan Lynne L

Protein science : a publication of the Protein Society 20230201 2

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 ...[more]

PMID: 36585831

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