Research Papers: Sensing

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

[+] Author Affiliations
Vinh Nguyen Du Le

McMaster University, Department of Medical Physics and Applied Radiation Sciences, Hamilton, Ontario L8S 4L8, Canada

Michael S. Patterson, Thomas J. Farrell, Joseph E. Hayward

McMaster University, Department of Medical Physics and Applied Radiation Sciences, Hamilton, Ontario L8S 4L8, Canada

Juravinski Cancer Centre, Hamilton, Ontario L8V 5C2, Canada

Qiyin Fang

McMaster University, Department of Engineering Physics, Hamilton, Ontario L8S 4L8, Canada

J. Biomed. Opt. 20(12), 127003 (Dec 22, 2015). doi:10.1117/1.JBO.20.12.127003
History: Received August 1, 2015; Accepted November 12, 2015
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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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© 2015 Society of Photo-Optical Instrumentation Engineers

Citation

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", J. Biomed. Opt. 20(12), 127003 (Dec 22, 2015). ; http://dx.doi.org/10.1117/1.JBO.20.12.127003


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