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Two-colour live-cell nanoscale imaging of intracellular targets.

ABSTRACT: Stimulated emission depletion (STED) nanoscopy allows observations of subcellular dynamics at the nanoscale. Applications have, however, been severely limited by the lack of a versatile STED-compatible two-colour labelling strategy for intracellular targets in living cells. Here we demonstrate a universal labelling method based on the organic, membrane-permeable dyes SiR and ATTO590 as Halo and SNAP substrates. SiR and ATTO590 constitute the first suitable dye pair for two-colour STED imaging in living cells below 50?nm resolution. We show applications with mitochondria, endoplasmic reticulum, plasma membrane and Golgi-localized proteins, and demonstrate continuous acquisition for up to 3?min at 2-s time resolution.

SUBMITTER: Bottanelli F

PROVIDER: S-EPMC4785223 | biostudies-literature | 2016 Mar

REPOSITORIES: biostudies-literature

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Two-colour live-cell nanoscale imaging of intracellular targets.

Bottanelli Francesca F Kromann Emil B EB Allgeyer Edward S ES Erdmann Roman S RS Wood Baguley Stephanie S Sirinakis George G Schepartz Alanna A Baddeley David D Toomre Derek K DK Rothman James E JE Bewersdorf Joerg J

Nature communications 20160304

Stimulated emission depletion (STED) nanoscopy allows observations of subcellular dynamics at the nanoscale. Applications have, however, been severely limited by the lack of a versatile STED-compatible two-colour labelling strategy for intracellular targets in living cells. Here we demonstrate a universal labelling method based on the organic, membrane-permeable dyes SiR and ATTO590 as Halo and SNAP substrates. SiR and ATTO590 constitute the first suitable dye pair for two-colour STED imaging in ...[more]

PMID: 26940217

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