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Optical Engineering • new

Fluorescence decay data analysis correcting for detector pulse pile-up at very high count rates
Author(s): Matthias Patting; Paja Reisch; Marcus Sackrow; Rhys Dowler; Marcelle Koenig; Michael Wahl
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Paper 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.

Paper Details

Date Published: 22 January 2018
PDF: 5 pages
Opt. Eng. 57(3) 031305 doi: 10.1117/1.OE.57.3.031305
Published in: Optical Engineering Volume 57, Issue 3
Show Author Affiliations
Matthias Patting, PicoQuant GmbH (Germany)
Paja Reisch, PicoQuant GmbH (Germany)
Marcus Sackrow, PicoQuant GmbH (Germany)
Rhys Dowler, PicoQuant GmbH (Germany)
Marcelle Koenig, PicoQuant GmbH (Germany)
Michael Wahl, PicoQuant GmbH (Germany)


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