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Proceedings Paper

Spectral unmixing of multi-color tissue specific in vivo fluorescence in mice
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Paper Abstract

Fluorescence Molecular Tomography (FMT) has emerged as a powerful tool for monitoring biological functions in vivo in small animals. It provides the means to determine volumetric images of fluorescent protein concentration by applying the principles of diffuse optical tomography. Using different probes tagged to different proteins or cells, different biological functions and pathways can be simultaneously imaged in the same subject. In this work we present a spectral unmixing algorithm capable of separating signal from different probes when combined with the tomographic imaging modality. We show results of two-color imaging when the algorithm is applied to separate fluorescence activity originating from phantoms containing two different fluorophores, namely CFSE and SNARF, with well separated emission spectra, as well as Dsred- and GFP-fused cells in F5-b10 transgenic mice in vivo. The same algorithm can furthermore be applied to tissue-specific spectroscopy data. Spectral analysis of a variety of organs from control, DsRed and GFP F5/B10 transgenic mice showed that fluorophore detection by optical systems is highly tissue-dependent. Spectral data collected from different organs can provide useful insight into experimental parameter optimisation (choice of filters, fluorophores, excitation wavelengths) and spectral unmixing can be applied to measure the tissue-dependency, thereby taking into account localized fluorophore efficiency. Summed up, tissue spectral unmixing can be used as criteria in choosing the most appropriate tissue targets as well as fluorescent markers for specific applications.

Paper Details

Date Published: 11 July 2007
PDF: 8 pages
Proc. SPIE 6626, Molecular Imaging, 662609 (11 July 2007); doi: 10.1117/12.728410
Show Author Affiliations
Giannis Zacharakis, Foundation for Research and Technology-Hellas (Greece)
Rosy Favicchio, Foundation for Research and Technology-Hellas (Greece)
Anikitos Garofalakis, Foundation for Research and Technology-Hellas (Greece)
Stylianos Psycharakis, Foundation for Research and Technology-Hellas (Greece)
Clio Mamalaki, Foundation for Research and Technology-Hellas (Greece)
Jorge Ripoll, Foundation for Research and Technology-Hellas (Greece)

Published in SPIE Proceedings Vol. 6626:
Molecular Imaging
Kai Licha; Vasilis Ntziachristos, Editor(s)

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