Counting Fluorescent dye molecules on DNA origami by means of photon statistics

• Verfasst von: Kurz, Anton [VerfasserIn]
• Verfasst von: Schmied, Jürgen J. [VerfasserIn]
• Verfasst von: Grußmayer, Kristin Stefanie [VerfasserIn]
• Verfasst von: Holzmeister, Phil [VerfasserIn]
• Verfasst von: Tinnefeld, Philip [VerfasserIn]
• Verfasst von: Herten, Dirk-Peter [VerfasserIn]
• Titel: Counting Fluorescent dye molecules on DNA origami by means of photon statistics
• Verf.angabe: Anton Kurz, Jürgen J. Schmied, Kristin S. Grußmayer, Phil Holzmeister, Philip Tinnefeld, and Dirk-Peter Herten
• E-Jahr: 2013
• Jahr: 24 June 2013
• Umfang: 8 S.
• Teil: volume:9
• Teil: year:2013
• Teil: number:23
• Teil: month:12
• Teil: pages:4061-4068
• Teil: extent:8
• Fussnoten: Gesehen am 22.07.2021
• Titel Quelle: Enthalten in: Small
• Ort Quelle: Weinheim : Wiley-VCH, 2005
• Jahr Quelle: 2013
• Band/Heft Quelle: 9(2013), 23 vom: Dez., Seite 4061-4068
• ISSN Quelle: 1613-6829
• DOI: doi:10.1002/smll.201300619
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: DNA origami
• Sach-SW: photon antibunching
• Sach-SW: photon statistics
• Sach-SW: single molecule fluorescence spectroscopy
• K10plus-PPN: 1764148916

Abstract

Obtaining quantitative information about molecular assemblies with high spatial and temporal resolution is a challenging task in fluorescence microscopy. Single-molecule techniques build on the ability to count molecules one by one. Here, a method is presented that extends recent approaches to analyze the statistics of coincidently emitted photons to enable reliable counting of molecules in the range of 1-20. This method does not require photochemistry such as blinking or bleaching. DNA origami structures are labeled with up to 36 dye molecules as a new evaluation tool to characterize this counting by a photon statistics approach. Labeled DNA origami has a well-defined labeling stoichiometry and ensures equal brightness for all dyes incorporated. Bias and precision of the estimating algorithm are determined, along with the minimal acquisition time required for robust estimation. Complexes containing up to 18 molecules can be investigated non-invasively within 150 ms. The method might become a quantifying add-on for confocal microscopes and could be especially powerful in combination with STED/RESOLFT-type microscopy.