Self-quenched Fluorophore Dimers for DNA-PAINT and STED Microscopy.
Angew Chem Int Ed Engl
; 62(39): e202307538, 2023 09 25.
Article
em En
| MEDLINE
| ID: mdl-37581373
ABSTRACT
Super-resolution techniques like single-molecule localisation microscopy (SMLM) and stimulated emission depletion (STED) microscopy have been extended by the use of non-covalent, weak affinity-based transient labelling systems. DNA-based hybrid systems are a prominent example among these transient labelling systems, offering excellent opportunities for multi-target fluorescence imaging. However, these techniques suffer from higher background relative to covalently bound fluorophores, originating from unbound fluorophore-labelled single-stranded oligonucleotides. Here, we introduce short-distance self-quenching in fluorophore dimers as an efficient mechanism to reduce background fluorescence signal, while at the same time increasing the photon budget in the bound state by almost 2-fold. We characterise the optical and thermodynamic properties of fluorophore-dimer single-stranded DNA, and show super-resolution imaging applications with STED and SMLM with increased spatial resolution and reduced background.
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Base de dados:
MEDLINE
Assunto principal:
DNA
/
Imagem Individual de Molécula
Idioma:
En
Ano de publicação:
2023
Tipo de documento:
Article