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Picosecond wide-field time-correlated single photon counting fluorescence microscopy with a delay line anode detector

Research output: Contribution to journalArticle

Liisa Maija Hirvonen, Wolfgang Becker, James Milnes, Thomas Conneely, Stefan Smietana, Alix Marie Le Marois, Ottmar Jagutzki, Klaus Suhling

Original languageEnglish
Article number071101
Number of pages4
Issue number7
Accepted/In press3 Aug 2016
Published15 Aug 2016


  • Picosecond wide-field time_HIRVONEN_Accepted 3Aug2016_GOLD VoR

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    © 2016 Author(s). All article content, except where otherwise noted, is licensed under a Creative Commons Attribution (CC BY) license (

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We perform wide-field time-correlated single photon counting-based fluorescence lifetime imaging (FLIM) with a crossed delay line anode image intensifier, where the pulse propagation time yields the photon position. This microchannel plate-based detector was read out with conventional fast timing electronics and mounted on a fluorescence microscope with total internal reflection (TIR) illumination. The picosecond time resolution of this detection system combines low illumination intensity of microwatts with wide-field data collection. This is ideal for fluorescence lifetime imaging of cell membranes using TIR. We show that fluorescence lifetime images of living HeLa cells stained with membrane dye di-4-ANEPPDHQ exhibit a reduced lifetime near the coverslip in TIR compared to epifluorescence FLIM.

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