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2 March 2022 Optimal conditions for the phasor plot analysis of fluorescence lifetime data
Wonsang Hwang, Dongeun Kim, Dugyoung Kim
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Proceedings Volume PC11966, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIX; PC1196603 (2022) https://doi.org/10.1117/12.2608466
Event: SPIE BiOS, 2022, San Francisco, California, United States
Abstract
Phasor domain clustering of Fluorescence Lifetime data has gained a lot of attention because of fit-free algorithm and the ability to resolve heterogeneity of the sample. Despite the popularity, the optimal modulation frequency has not been discussed yet. Especially, an optimal condition must be considered where the two clusters are too close to discriminate, or it is challenging to acquire enough photons. In this study, we found optimal modulation frequency to resolve two clusters in the phasor domain. We theoretically derived the Davies-Bouldin index (DBI) of two clusters in the phasor domain and predicted the optimal frequency based on it. We verified the theoretical prediction via two different fluorescence droplets experiments.
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Wonsang Hwang, Dongeun Kim, and Dugyoung Kim "Optimal conditions for the phasor plot analysis of fluorescence lifetime data", Proc. SPIE PC11966, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIX, PC1196603 (2 March 2022); https://doi.org/10.1117/12.2608466
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KEYWORDS
Luminescence
Modulation
Photons
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