title: Experimental recovery of intrinsic fluorescence and fluorophore concentration in the presence of hemoglobin: spectral effect of scattering and absorption on fluorescence.
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).
authors.affiliation: McMaster University, Department of Medical Physics and Applied Radiation Sciences, Hamilton, Ontario L8S 4L8, Canada.
authors.forename: Vinh Nguyen
authors.initials: VN
authors.lastname: Du Le
authors.affiliation: McMaster University, Department of Medical Physics and Applied Radiation Sciences, Hamilton, Ontario L8S 4L8, CanadabJuravinski Cancer Centre, Hamilton, Ontario L8V 5C2, Canada.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Patterson
authors.affiliation: McMaster University, Department of Medical Physics and Applied Radiation Sciences, Hamilton, Ontario L8S 4L8, CanadabJuravinski Cancer Centre, Hamilton, Ontario L8V 5C2, Canada.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Farrell
authors.affiliation: McMaster University, Department of Medical Physics and Applied Radiation Sciences, Hamilton, Ontario L8S 4L8, CanadabJuravinski Cancer Centre, Hamilton, Ontario L8V 5C2, Canada.
authors.forename: Joseph E
authors.initials: JE
authors.lastname: Hayward
authors.affiliation: McMaster University, Department of Engineering Physics, Hamilton, Ontario L8S 4L8, Canada.
authors.forename: Qiyin
authors.initials: Q
authors.lastname: Fang
mesh_terms: D002244:Carbon; D003198:Computer Simulation; D019793:Fluorescein / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D006454:Hemoglobins / Q000737:chemistry*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008863:Microspheres; D009092:Mucous Membrane / Q000473:pathology; D019047:Phantoms, Imaging; D011137:Polystyrenes / Q000737:chemistry; D011336:Probability; D015203:Reproducibility of Results; D012542:Scattering, Radiation; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013053:Spectrophotometry
chemical_list: D005456:Fluorescent Dyes; D006454:Hemoglobins; D011137:Polystyrenes; C028433:chinese ink; D002244:Carbon; D019793:Fluorescein