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Sub-μs time resolution in wide-field time-correlated single photon counting microscopy obtained from the photon event phosphor decay

Hirvonen, Liisa M.; Petrášek, Zdeněk; Beeby, Andrew; Suhling, Klaus

Sub-μs time resolution in wide-field time-correlated single photon counting microscopy obtained from the photon event phosphor decay Thumbnail


Authors

Liisa M. Hirvonen

Zdeněk Petrášek

Klaus Suhling



Abstract

Fast frame rate complementary metal–oxide–semiconductor cameras in combination with photon counting image intensifiers can be used for microsecond resolution wide-field fluorescence lifetime imaging with single photon sensitivity, but the time resolution is limited by the camera exposure time. We show here how the image intensifier's P20 phosphor afterglow can be exploited for accurate timing of photon arrival well below the camera exposure time. By taking ratios of the intensity of the photon events in two subsequent frames, photon arrival times were determined with 300 ns precision with 18.5 μs frame exposure time (54 kHz camera frame rate). Decays of ruthenium and iridium-containing compounds with around 1 μs lifetimes were mapped with this technique, including in living HeLa cells, using excitation powers below 0.5 μW. Details of the implementation to calculate the arrival time from the photon event intensity ratio are discussed, and we speculate that by using an image intensifier with a faster phosphor decay to match a higher camera frame rate, photon arrival time measurements on the nanosecond time scale could be possible.

Citation

Hirvonen, L. M., Petrášek, Z., Beeby, A., & Suhling, K. (2015). Sub-μs time resolution in wide-field time-correlated single photon counting microscopy obtained from the photon event phosphor decay. New Journal of Physics, 17(2), https://doi.org/10.1088/1367-2630/17/2/023032

Journal Article Type Article
Acceptance Date Jan 9, 2015
Publication Date Feb 10, 2015
Deposit Date May 7, 2015
Publicly Available Date Mar 29, 2024
Journal New Journal of Physics
Publisher IOP Publishing
Peer Reviewed Peer Reviewed
Volume 17
Issue 2
DOI https://doi.org/10.1088/1367-2630/17/2/023032

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Publisher Licence URL
http://creativecommons.org/licenses/by/4.0/

Copyright Statement
Content from this work may be used under the terms of the Creative Commons Attribution 3.0 licence. Any further distribution of this work must maintain attribution to the author(s) and the title of the work, journal citation and DOI.





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