Fluorescence lifetime spectroscopy in multiple-scattering environments: an application to biotechnology

Albert E. Cerussi; Enrico Gratton; Sergio Fantini

doi:10.1117/12.351028

2 July 1999 Fluorescence lifetime spectroscopy in multiple-scattering environments: an application to biotechnology

Albert E. Cerussi, Enrico Gratton, Sergio Fantini

Proceedings Volume 3600, Biomedical Imaging: Reporters, Dyes, and Instrumentation; (1999) https://doi.org/10.1117/12.351028
Event: BiOS '99 International Biomedical Optics Symposium, 1999, San Jose, CA, United States

Abstract

Over the past few years, there has been significant research activity devoted to the application of fluorescence spectroscopy to strongly scattering media, where photons propagate diffusely. Much of this activity focused on fluorescence as a source of contrast enhancement in optical tomography. Our efforts have emphasized the quantitative recovery of fluorescence parameters for spectroscopy. Using a frequency-domain diffusion-based model, we have successfully recovered the lifetime, the absolute quantum yield, the fluorophore concentration, and the emission spectrum of the fluorophore, as well as the absorption and the reduced scattering coefficients at the emission wavelength of the medium in different measurements. In this contribution, we present a sensitive monitor of the binding between ethidium bromide and bovine cells in fresh milk. The spectroscopic contrast was the approximately tenfold increase in the ethidium bromide lifetime upon binding to DNA. The measurement clearly demonstrated that we could quantitatively measure the density of cells in the milk, which is an application vital to the tremendous economic burden of bovine subclinical mastitis detection. Furthermore, we may in principle use the spirit of this technique as a quantitative monitor of the binding of fluorescent drugs inside tissues. This is a first step towards lifetime spectroscopy in tissues.

Citation Download Citation

Albert E. Cerussi, Enrico Gratton, and Sergio Fantini "Fluorescence lifetime spectroscopy in multiple-scattering environments: an application to biotechnology", Proc. SPIE 3600, Biomedical Imaging: Reporters, Dyes, and Instrumentation, (2 July 1999); https://doi.org/10.1117/12.351028

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