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

Published version
Peer-reviewed

Type

Article

Change log

Authors

Bottanelli, Francesca 
Kromann, Emil B 
Allgeyer, Edward S 
Erdmann, Roman S 
Wood Baguley, Stephanie 

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.

Description

Keywords

Animals, COS Cells, Chlorocebus aethiops, HeLa Cells, Heterocyclic Compounds, 4 or More Rings, Humans, Luminescent Proteins, Microscopy, Fluorescence, Nanotechnology, Rhodamines

Journal Title

Nat Commun

Conference Name

Journal ISSN

2041-1723
2041-1723

Volume Title

7

Publisher

Springer Science and Business Media LLC
Sponsorship
Wellcome Trust (092096/Z/10/Z)
Cancer Research Uk (None)