Abstract
Although conventional autofluorescence spectroscopy, in which fluorescence emission spectra are recorded for fixed excitation wavelengths, has demonstrated good performance in tissue diagnosis, it suffers from prolonged data acquisition time and broad-band fluorescence features. Synchronous spectroscopy has been proposed to overcome the limitations of conventional fluorescence spectroscopy but has not been applied to imaging for tissue diagnosis in vivo. Our group has developed a synchronous fluorescence imaging system to combine the great diagnostic potential of synchronous spectroscopy and the large field of view of imaging for cancer diagnosis. This system has been tested in a mouse skin model to capture synchronous fluorescence images. A simple discriminant analysis method and a more complicated multi-variate statistical method have been developed to generate a single diagnostic image from a large number of raw fluorescence images. Moreover, it was demonstrated that the diagnostic image generated from synchronous data is comparable to that generated from full spectral data in classification accuracy.
Highlights
IntroductionConventional fluorescence spectroscopy uses either a fixed-wavelength excitation (λexc) to produce an emission spectrum or a fixed wavelength emission (λemm) to record an excitation spectrum
Tissue autofluorescence may be contributed by several endogenous fluorophores such as aromatic amino acids, structural proteins, nicotiamide adenine dinucleotide (NADH), flavin adenine dinucleotide (FAD), porphyrins, lipopigments, and other biological components.Conventional fluorescence spectroscopy uses either a fixed-wavelength excitation to produce an emission spectrum or a fixed wavelength emission to record an excitation spectrum
A vast majority of information in the raw data has been taken advantage of. This approach yields a more accurate diagnostic image than the raw data set because some fractions of the raw data that don’t exhibit significant difference between malignant and normal regions have been removed
Summary
Conventional fluorescence spectroscopy uses either a fixed-wavelength excitation (λexc) to produce an emission spectrum or a fixed wavelength emission (λemm) to record an excitation spectrum. Changes in fluorescence profiles using fixed excitation/emission have been reported by these researchers, the changes do not provide "unique narrow-band spectral signatures" useful for unequivocal diagnostic purposes, which did not fully exploit the diagnostic potential of fluorescence spectroscopy. This method requires prolonged acquisition time to collect full spectral data. The fluorescence signal is recorded while both λemm and λexc are simultaneously scanned. The intensity of the synchronous signal Is, can be written as the product of three functions as follows 5,6:
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