Saturation-compensated measurements for fluorescence lifetime imaging microscopy

Abstract

Fluorophore saturation is the key factor limiting the speed and excitation range of fluorescence lifetime imaging microscopy (FLIM). For example, fluorophore saturation causes incorrect lifetime measurements when using conventional frequency-domain FLIM at high excitation powers. In this Letter, we present an analytical theoretical description of this error and present a method for compensating for this error in order to extract correct lifetime measurements in the limit of fluorophore saturation. We perform a series of simulations and experiments to validate our methods. The simulations and experiments show a 13.2× and a 2.6× increase in excitation range, respectively. The presented method is based on algorithms that can be easily applied to existing FLIM setups.

Publication
Optics Letters, vol. 42, no. 1, pp. 155-158
Yide Zhang
Yide Zhang
NIH K99 Postdoctoral Fellow

My research is interdisciplinary and focused on developing new types of optical imaging techniques that could advance the work of other researchers and medical personnel in a wide variety of fields. Currently, I am developing next-generation photoacoustic and ultrafast imaging techniques that can observe biological and physical phenomena that are too fast to be imaged with existing methods. The observation of the ultrafast phenomena could provide a better understanding of the fundamentals of life and physical sciences. I am also developing novel quantum imaging approaches that can investigate biological organisms with an imaging performance that cannot be achieved using classical optical imaging. In my free time, I enjoy cooking, hiking, cycling, and traveling.

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