Work

Optimal modulation frequency of phasor-domain clustering and analysis for fluorescence lifetime data

Year: 2026

Type: article

Abstract: We present a mathematical framework for determining the optimal modulation frequency for phasor-domain clustering using the Davies–Bouldin index (DBI). By analytically formulating the DBI for two clusters, we model the relationship between photon counts, signal-to-noise ratio (SNR), and cluster dispersion to identify the frequency that maximizes separation. Our model predicts a non-monotonic dependence of the optimal frequency on SNR, with f opt tending toward zero in the high-SNR limit as the contribution of dark counts becomes negligible. These theoretical findings were experimentally validated using fluorescent droplets and biological samples with distinct lifetimes, demonstrating that crosstalk pixel trends align with the predicted DBI.

Source: Optics Express

DOI: 10.1364/oe.601428

Authors: Wonsang Hwang, Dongeun Kim, Sinyoung Jeong, Iván Coto Hernández, Lukas-Matthaeus Altenburger, Dug Young Kim