Fluorescence decay data analysis correcting for detector pulse pile-up at very high count rates

Matthias Patting; Paja Reisch; Marcus Sackrow; Rhys Dowler; Marcelle Koenig; Michael Wahl

doi:10.1117/1.OE.57.3.031305

22 January 2018 Fluorescence decay data analysis correcting for detector pulse pile-up at very high count rates

Matthias Patting, Paja Reisch, Marcus Sackrow, Rhys Dowler, Marcelle Koenig, Michael Wahl

Author Affiliations +

Funded by: German Federal Ministry of Education and Research (Bundesministerium für Bildung und Forschung)

Optical Engineering, Vol. 57, Issue 3, 031305 (January 2018). https://doi.org/10.1117/1.OE.57.3.031305

Abstract

Using time-correlated single photon counting for the purpose of fluorescence lifetime measurements is usually limited in speed due to pile-up. With modern instrumentation, this limitation can be lifted significantly, but some artifacts due to frequent merging of closely spaced detector pulses (detector pulse pile-up) remain an issue to be addressed. We propose a data analysis method correcting for this type of artifact and the resulting systematic errors. It physically models the photon losses due to detector pulse pile-up and incorporates the loss in the decay fit model employed to obtain fluorescence lifetimes and relative amplitudes of the decay components. Comparison of results with and without this correction shows a significant reduction of systematic errors at count rates approaching the excitation rate. This allows quantitatively accurate fluorescence lifetime imaging at very high frame rates.

Citation Download Citation

Matthias Patting, Paja Reisch, Marcus Sackrow, Rhys Dowler, Marcelle Koenig, and Michael Wahl "Fluorescence decay data analysis correcting for detector pulse pile-up at very high count rates," Optical Engineering 57(3), 031305 (22 January 2018). https://doi.org/10.1117/1.OE.57.3.031305

Received: 2 October 2017; Accepted: 20 December 2017; Published: 22 January 2018

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