Extraction of intrinsic fluorescence from single fiber fluorescence measurements on a turbid medium: experimental validation

Ute Gamm, Chris Hoy, Floor Zaane, Dick Sterenborg, Chad Kanick, Dominic Robinson, Arjen Amelink

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The detailed mechanisms associated with the influence of scattering and absorption properties on the fluorescence intensity sampled by a single optical fiber have recently been elucidated based on Monte Carlo simulated data. Here we develop an experimental single fiber fluorescence (SFF) spectroscopy setup and validate the Monte Carlo data and semi-empirical model equation that describes the SFF signal as a function of scattering. We present a calibration procedure that corrects the SFF signal for all system- related, wavelength dependent transmission efficiencies to yield an absolute value of intrinsic fluorescence. The validity of the Monte Carlo data and semi- empirical model is demonstrated using a set of fluorescent phantoms with varying concentrations of Intralipid to vary the scattering properties, yielding a wide range of reduced scattering coefficients (mu'(s) = 0-7 mm(-1)). We also introduce a small modification to the model to account for the case of mu's = 0 mm(-1) and show its relation to the experimental, simulated and theoretically calculated value of SFF intensity in the absence of scattering. Finally, we show that our method is also accurate in the presence of absorbers by performing measurements on phantoms containing red blood cells and correcting for their absorption properties. (C) 2014 Optical Society of America
Original languageUndefined/Unknown
Pages (from-to)1913-1925
Number of pages13
JournalBiomedical Optics Express
Issue number6
Publication statusPublished - 2014

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