Two-colour live-cell nanoscale imaging of intracellular targets.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 26940217.
- Also identified by DOI 10.1038/ncomms10778 and PMC identifier 4785223.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
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.
Medical subject headings
- Heterocyclic Compounds, 4 or More Rings
- Luminescent Proteins
- Microscopy, Fluorescence
- Nanotechnology
- Rhodamines