Two-colour live-cell nanoscale imaging of intracellular targets

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 uni...

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Detalles Bibliográficos
Autores principales: Bottanelli, Francesca, Kromann, Emil B., Allgeyer, Edward S., Erdmann, Roman S., Wood Baguley, Stephanie, Sirinakis, George, Schepartz, Alanna, Baddeley, David, Toomre, Derek K., Rothman, James E., Bewersdorf, Joerg
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4785223/
https://www.ncbi.nlm.nih.gov/pubmed/26940217
http://dx.doi.org/10.1038/ncomms10778
Descripción
Sumario: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.

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