Document Type

Article

Article Version

Publisher's PDF

Publication Date

2017

Abstract

We report chemometric wide-field fluorescence microscopy for imaging the spatial distribution and concentration of endogenous fluorophores in thin tissue sections. Nonnegative factorization aided by spatial diversity is used to learn both the spectral signature and the spatial distribution of endogenous fluorophores from microscopic fluorescence color images obtained under broadband excitation and detection. The absolute concentration map of individual fluorophores is derived by comparing the fluorescence from “pure” fluorophores under the identical imaging condition following the identification of the fluorescence species by its spectral signature. This method is then demonstrated by characterizing the concentration map of endogenous fluorophores (including tryptophan, elastin, nicotinamide adenine dinucleotide, and flavin adenine dinucleotide) for lung tissue specimens. The absolute concentrations of these fluorophores are all found to decrease significantly from normal, perilesional, to cancerous (squamous cell carcinoma) tissue. Discriminating tissue types using the absolute fluorophore concentration is found to be significantly more accurate than that achievable with the relative fluorescence strength. Quantification of fluorophores in terms of the absolute concentration map is also advantageous in eliminating the uncertainties due to system responses or measurement details, yielding more biologically relevant data, and simplifying the assessment of competing imaging approaches.

Comments

Copyright 2017 Society of Photo-optical Instrumentation Engineers (SPIE)

The final publisher PDF has been archived here with permission from the copyright holder.

Publication Title

Journal of biomedical optics

Published Citation

Xu, Z., Reilley, M., Li, R., & Xu, M. (2017). Mapping absolute tissue endogenous fluorophore concentrations with chemometric wide-field fluorescence microscopy. Journal of biomedical optics, 22(6), 066009. doi: 10.1117/1.JBO.22.6.066009.

DOI

10.1117/1.JBO.22.6.066009

Peer Reviewed

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