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Updated: Jun 24, 2026

Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy
Published on: January 18, 2017
Instrument response standard in time-resolved fluorescence
Rafal Luchowski1, Zygmunt Gryczynski, Pabak Sarkar
1Department of Molecular Biology and Immunology, Center for Commercialization of Fluorescence Technologies, UNTHSC, Fort Worth, Texas 76107, USA.
Abstract:
The fluorescence of LDS 798 dye in aqueous solution has a very short lifetime of 24 ps, independent of excitation wavelength. The time response of common photon counting detectors depends on the wavelength of the registered photon. In lifetime measurements, the instrument response function (IRF) is usually approximated by the temporal profile of the scattered excitation light. Because lambda(Exc) is typically much shorter than lambda(Em), a systematic error may be present in these measurements. We demonstrate that the fluorescence decay of LDS 798 is a better approximation of IRF, in particular, for avalanche photodiodes used in the near infrared spectral region.
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