Abstract
Fluorescence lifetime imaging (FLIM) has previously been shown to provide contrast between normal and diseased tissue. Here we present progress towards clinical and preclinical FLIM endoscopy of tissue autofluorescence, demonstrating a flexible wide‐field endoscope that utilised a low average power blue picosecond laser diode excitation source and was able to acquire ∼mm‐scale spatial maps of autofluorescence lifetimes from fresh ex vivo diseased human larynx biopsies in ∼8 seconds using an average excitation power of ∼0.5 mW at the specimen. To illustrate its potential for FLIM at higher acquisition rates, a higher power mode‐locked frequency doubled Ti:Sapphire laser was used to demonstrate FLIM of ex vivo mouse bowel at up to 2.5 Hz using 10 mW of average excitation power at the specimen. (© 2014 WILEY‐VCH Verlag GmbH &Co. KGaA, Weinheim)
Highlights
Fluorescence lifetime imaging (FLIM) [1, 2] of tissue autofluorescence has been shown to provide labelfree contrast between different types and states of tissue that may help differentiate between normal and diseased human tissue, e.g. [3,4,5,6,7,8]
Having established that we can utilise a single multimode excitation fibre to efficiently illuminate a 3 mm field of view (FOV), we studied the feasibility of using low cost gain-switched picosecond diodes for FLIM of tissue autofluorescence
We have demonstrated the feasibility of flexible wide-field FLIM endoscopy utilising low average power blue excitation sources applied to tissue autofluorescence
Summary
Fluorescence lifetime imaging (FLIM) [1, 2] of tissue autofluorescence has been shown to provide labelfree contrast between different types and states of tissue that may help differentiate between normal and diseased human tissue, e.g. [3,4,5,6,7,8]. Fluorescence lifetime imaging (FLIM) [1, 2] of tissue autofluorescence has been shown to provide labelfree contrast between different types and states of tissue that may help differentiate between normal and diseased human tissue, e.g. Building on the pioneering single-point clinical measurements of autofluorescence lifetime, e.g. [9, 10] and subsequent work recently reviewed in 2012 [11], clinical FLIM can combine lifetime contrast with morphological information to provide a direct comparison between different spatial regions – making it easier to spot differences with respect to “normal” tissue, e.g. for diagnostic screening and potentially enabling margins of diseased tissue to be identified. There have been relatively few clinical FLIM studies, partly due to a lack of suitable instrumentation.
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