Blind and reference-free fluorescence lifetime estimation via consumer time-of-flight sensors

Fluorescence lifetime imaging (FLI) is a popular method for extracting useful information that is otherwise unavailable from a conventional intensity image. Usually, however, it requires expensive equipment, is often limited to either distinctly frequency- or time-domain modalities, and demands calibration measurements and precise knowledge of the illumination signal. Here, we present a generalized time-based, cost-effective method for estimating lifetimes by repurposing a consumer-grade time-of-flight sensor. By developing mathematical theory that unifies time- and frequency-domain approaches, we can interpret a time-based signal as a combination of multiple frequency measurements. We show that we can estimate lifetimes without knowledge of the illumination signal and without any calibration. We experimentally demonstrate this blind, reference-free method using a quantum dot solution and discuss the method’s implementation in FLI applications.