An extended scheme for counting fluorescent molecules by photon-antibunching
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 singl...
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| Main Authors: | , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
2010
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| In: |
Laser physics
Year: 2010, Volume: 20, Issue: 1, Pages: 119-124 |
| ISSN: | 1555-6611 |
| DOI: | 10.1134/S1054660X09170204 |
| Online Access: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1134/S1054660X09170204
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| Author Notes: | Haisen Ta, Jürgen Wolfrum & Dirk-Peter Herten |
MARC
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| 520 | |a 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. | ||
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