Abstract
There is an increasing interest in the application of fluorescence lifetime imaging (FLIM) for medical diagnosis. Central to the clinical translation of FLIM technology is the development of compact and high-speed clinically compatible systems. We present a handheld probe design consisting of a small maneuverable box fitted with a rigid endoscope, capable of continuous lifetime imaging at multiple emission bands simultaneously. The system was characterized using standard fluorescent dyes. The performance was then further demonstrated by imaging a hamster cheek pouch in vivo, and oral mucosa tissue both ex vivo and in vivo, all using safe and permissible exposure levels. Such a design can greatly facilitate the evaluation of FLIM for oral cancer imaging in vivo.
Highlights
In fluorescence lifetime imaging (FLIM), the fluorescence lifetime is measured at each spatially resolvable location within a fluorescence image [1]
We report the first demonstration of real-time deconvolution of the instrument response from the fluorescence decay at each pixel of the image, which allowed accurate real-time lifetime map estimation and visualization at multiple spectral bands simultaneously
For the 25 um excitation fiber and the 50 mm focal length objective, the full width at half maximum (FWHM) of the point spread function (PSF) was calculated to be 44.8 um, which is in close agreement with the corresponding qualitatively value determined by imaging the USAF resolution target as
Summary
In fluorescence lifetime imaging (FLIM), the fluorescence lifetime is measured at each spatially resolvable location within a fluorescence image [1]. There has been an increasing interest in evaluating the application of endogenous FLIM for clinical diagnosis. The FLIM endoscope designs proposed far, including those summarized below, are still not well suited for clinical applications. The fluorescence lifetime can be implemented in the time domain by directly measuring the fluorescence decay following pulsed light excitation [1]. Wide-field time domain FLIM can be implemented using time-gated imaging. Point scanning time domain FLIM can be implemented using either time-correlated single-photon counting (TCSPC) or the direct pulse recording approach. The fluorescence intensity decay is measured as function of time upon a single excitation pulse using high-bandwidth detectors and digitizers
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