Experimental recovery of intrinsic fluorescence and fluorophore concentration in the presence of hemoglobin: spectral effect of scattering and absorption on fluorescence

Vinh Nguyen Du Le; Michael S. Patterson; Thomas J. Farrell; Joseph E. Hayward; Qiyin Fang

doi:10.1117/1.JBO.20.12.127003

22 December 2015 Experimental recovery of intrinsic fluorescence and fluorophore concentration in the presence of hemoglobin: spectral effect of scattering and absorption on fluorescence

Vinh Nguyen Du Le, Michael S. Patterson, Thomas J. Farrell, Joseph E. Hayward, Qiyin Fang

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Journal of Biomedical Optics, Vol. 20, Issue 12, 127003 (December 2015). https://doi.org/10.1117/1.JBO.20.12.127003

Abstract

The ability to recover the intrinsic fluorescence of biological fluorophores is crucial to accurately identify the fluorophores and quantify their concentrations in the media. Although some studies have successfully retrieved the fluorescence spectral shape of known fluorophores, the techniques usually came with heavy computation costs and did not apply for strongly absorptive media, and the intrinsic fluorescence intensity and fluorophore concentration were not recovered. In this communication, an experimental approach was presented to recover intrinsic fluorescence and concentration of fluorescein in the presence of hemoglobin (Hb). The results indicated that the method was efficient in recovering the intrinsic fluorescence peak and fluorophore concentration with an error of 3% and 10%, respectively. The results also suggested that chromophores with irregular absorption spectra (e.g., Hb) have more profound effects on fluorescence spectral shape than chromophores with monotonic absorption and scattering spectra (e.g., black India ink and polystyrene microspheres).

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Vinh Nguyen Du Le, Michael S. Patterson, Thomas J. Farrell, Joseph E. Hayward, and Qiyin Fang "Experimental recovery of intrinsic fluorescence and fluorophore concentration in the presence of hemoglobin: spectral effect of scattering and absorption on fluorescence," Journal of Biomedical Optics 20(12), 127003 (22 December 2015). https://doi.org/10.1117/1.JBO.20.12.127003

Published: 22 December 2015

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