Segmented Frequency-domain Fluorescence Lifetime Measurements: Minimizing the Effects of Photobleaching Within a Multi-component System |
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Authors: | Hadi M Marwani Mark Lowry Patrick Keating Isiah M Warner Robert L Cook |
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Institution: | (1) Department of Chemistry, Louisiana State University, Baton Rouge, LA 70803, USA;(2) Department of Chemistry, Southern University at Baton Rouge, Baton Rouge, LA 70813, USA |
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Abstract: | This study introduces a newly developed frequency segmentation and recombination method for frequency-domain fluorescence
lifetime measurements to address the effects of changing fractional contributions over time and minimize the effects of photobleaching
within multi-component systems. Frequency segmentation and recombination experiments were evaluated using a two component
system consisting of fluorescein and rhodamine B. Comparison of experimental data collected in traditional and segmented fashion
with simulated data, generated using different changing fractional contributions, demonstrated the validity of the technique.
Frequency segmentation and recombination was also applied to a more complex system consisting of pyrene with Suwannee River
fulvic acid reference and was shown to improve recovered lifetimes and fractional intensity contributions. It was observed
that photobleaching in both systems led to errors in recovered lifetimes which can complicate the interpretation of lifetime
results. Results showed clear evidence that the frequency segmentation and recombination method reduced errors resulting from
a changing fractional contribution in a multi-component system, and allowed photobleaching issues to be addressed by commercially
available instrumentation.
Electronic supplementary material The online version of this article doi: contains supplementary material, which is available to authorized users. |
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Keywords: | Time-resolved fluorescence Frequency-domain lifetime measurements Frequency segmentation Photobleaching Multi-component system Data analysis |
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