An extended scheme for counting fluorescent molecules by photon-antibunching

• Verfasst von: Ta, Haisen [VerfasserIn]
• Verfasst von: Wolfrum, Jürgen [VerfasserIn]
• Verfasst von: Herten, Dirk-Peter [VerfasserIn]
• Titel: An extended scheme for counting fluorescent molecules by photon-antibunching
• Verf.angabe: Haisen Ta, Jürgen Wolfrum & Dirk-Peter Herten
• Jahr: 2010
• Umfang: 6 S.
• Fussnoten: Erstmals am 29 July 2009 online veröffentlicht ; Gesehen am 13.09.2023
• Titel Quelle: Enthalten in: Laser physics
• Ort Quelle: Bristol [u.a.] : IOP Publ., 2006
• Jahr Quelle: 2010
• Band/Heft Quelle: 20(2010), 1, Seite 119-124
• ISSN Quelle: 1555-6611
• DOI: doi:10.1134/S1054660X09170204
• Datenträger: Online-Ressource
• Sprache: eng
• Bibliogr. Hinweis: Erscheint auch als : Druck-Ausgabe: Ta, Haisen: An extended scheme for counting fluorescent molecules by photon-antibunching. - 2010
• Sach-SW: Dichroic Mirror
• Sach-SW: Forster Resonance Energy Transfer
• Sach-SW: Observation Volume
• Sach-SW: Single Molecule
• Sach-SW: Time Correlate Single Photon Counting
• K10plus-PPN: 1859418732

Abstract

Acquisition of quantitative information from microscopic biological samples is highly desirable in the context of the emerging field of systems biology. We derive a statistical approach to estimate the number of fluorescent molecules in the observation volume based on a confocal microscope for single-molecule detection. The method employs ps-pulsed laser sources for excitation and time-correlated single-photon counting with 4 avalanche photon diodes (APDs) for detection of individual photons. The feasibility for estimating the number of molecules is shown based on simultaneous emission and detection of multiple photons (photon-antibunching) under realistic experimental conditions. In theory, it should be possible to estimate the number of molecules with errors of less than 1% by using novel photo-stabilizing agents. The proposed method puts into perspective its application for high-resolution microscopy without the need for photo-switching or photo-activation of fluorescence dyes.