1. Two-colour live-cell nanoscale imaging of intracellular targets
- Author
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Stephanie Wood Baguley, Edward S. Allgeyer, Alanna Schepartz, David Baddeley, George Sirinakis, Roman S. Erdmann, Emil B. Kromann, Francesca Bottanelli, James E. Rothman, Joerg Bewersdorf, Derek Toomre, Allgeyer, Edward [0000-0002-2187-4423], Sirinakis, George [0000-0002-4762-422X], and Apollo - University of Cambridge Repository
- Subjects
0301 basic medicine ,or More Rings ,Science ,General Physics and Astronomy ,Biology ,Heterocyclic Compounds, 4 or More Rings ,General Biochemistry, Genetics and Molecular Biology ,Fluorescence ,Article ,Cercopithecus aethiops ,03 medical and health sciences ,symbols.namesake ,Heterocyclic Compounds ,Chlorocebus aethiops ,2.1 Biological and endogenous factors ,Animals ,Humans ,Nanotechnology ,Aetiology ,Cytoskeleton ,Luminescent Proteins ,Microscopy ,FOS: Nanotechnology ,Multidisciplinary ,Rhodamines ,Endoplasmic reticulum ,Vesicle ,STED microscopy ,General Chemistry ,Golgi apparatus ,4 or More Rings ,Fusion protein ,Cell biology ,030104 developmental biology ,Microscopy, Fluorescence ,Hela Cells ,COS Cells ,Biophysics ,symbols ,Generic health relevance ,Intracellular ,HeLa Cells - 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., The intracellular applications of STED microscopy are limited by the availability of dyes. Here the authors develop a two-colour labelling strategy based on SiR and ATTO590 dyes, and apply their strategy to image various subcellular membrane compartments.
- Published
- 2016