Molecule counts in localization microscopy with organic fluorophores

• Verfasst von: Karathanasis, Christos [VerfasserIn]
• Verfasst von: Heilemann, Mike [VerfasserIn]
• Titel: Molecule counts in localization microscopy with organic fluorophores
• Verf.angabe: Christos Karathanasis, Franziska Fricke, Gerhard Hummer, and Mike Heilemann
• E-Jahr: 2017
• Jahr: April 19, 2017
• Umfang: 7 S.
• Teil: volume:18
• Teil: year:2017
• Teil: number:8
• Teil: pages:942-948
• Teil: extent:7
• Fussnoten: Gesehen am 16.10.2018 ; First published: 14 February 2017
• Titel Quelle: Enthalten in: ChemPhysChem
• Ort Quelle: Weinheim : Wiley-VCH Verl., 2000
• Jahr Quelle: 2017
• Band/Heft Quelle: 18(2017), 8, Seite 942-948
• ISSN Quelle: 1439-7641
• DOI: doi:10.1002/cphc.201601425
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: molecular counting
• Sach-SW: organic fluorophores
• Sach-SW: photokinetic analysis
• Sach-SW: single-molecule imaging
• Sach-SW: super-resolution microscopy
• K10plus-PPN: 1581945795

Abstract

Single-molecule localization microscopy (SMLM) can be used to count fluorescently labeled molecules even when they are not individually resolved. We demonstrate SMLM molecule counting for nucleic acids labeled with the organic fluorophore Alexa Fluor 647 and imaged under photoswitching conditions. From the observed distributions of the number of fluorophore blinking events, we extract the number of fluorophores per spot using a statistical model. We validate the molecule counting method for single Alexa Fluor 647 fluorophores, and for trimers of Alexa Fluor 647 constructed on a DNA origami structure. This simple counting strategy enables quantitative super-resolution imaging with organic fluorophores.