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 +

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.

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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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