Du Le Vinh Nguyen, Patterson Michael S, Farrell Thomas J, Hayward Joseph E, Fang Qiyin
McMaster University, Department of Medical Physics and Applied Radiation Sciences, Hamilton, Ontario L8S 4L8, Canada.
McMaster University, Department of Medical Physics and Applied Radiation Sciences, Hamilton, Ontario L8S 4L8, CanadabJuravinski Cancer Centre, Hamilton, Ontario L8V 5C2, Canada.
J Biomed Opt. 2015;20(12):127003. doi: 10.1117/1.JBO.20.12.127003.
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).
恢复生物荧光团固有荧光的能力对于准确识别荧光团并量化其在介质中的浓度至关重要。尽管一些研究已成功恢复了已知荧光团的荧光光谱形状，但这些技术通常伴随着高昂的计算成本，且不适用于强吸收性介质，同时也未能恢复固有荧光强度和荧光团浓度。在本通讯中，提出了一种实验方法，用于在存在血红蛋白（Hb）的情况下恢复荧光素的固有荧光和浓度。结果表明，该方法在恢复固有荧光峰和荧光团浓度方面效率较高，误差分别为3%和10%。结果还表明，具有不规则吸收光谱的发色团（如Hb）对荧光光谱形状的影响比具有单调吸收和散射光谱的发色团（如黑色印度墨水和聚苯乙烯微球）更为显著。
