title: Quantitative performance characterization of three-dimensional noncontact fluorescence molecular tomography.
abstract: Fluorescent proteins and dyes are routine tools for biological research to describe the behavior of genes, proteins, and cells, as well as more complex physiological dynamics such as vessel permeability and pharmacokinetics. The use of these probes in whole body in vivo imaging would allow extending the range and scope of current biomedical applications and would be of great interest. In order to comply with a wide variety of application demands, in vivo imaging platform requirements span from wide spectral coverage to precise quantification capabilities. Fluorescence molecular tomography (FMT) detects and reconstructs in three dimensions the distribution of a fluorophore in vivo. Noncontact FMT allows fast scanning of an excitation source and noninvasive measurement of emitted fluorescent light using a virtual array detector operating in free space. Here, a rigorous process is defined that fully characterizes the performance of a custom-built horizontal noncontact FMT setup. Dynamic range, sensitivity, and quantitative accuracy across the visible spectrum were evaluated using fluorophores with emissions between 520 and 660 nm. These results demonstrate that high-performance quantitative three-dimensional visible light FMT allowed the detection of challenging mesenteric lymph nodes in vivo and the comparison of spectrally distinct fluorescent reporters in cell culture.
authors.affiliation: Imperial College London, Hammersmith Hospital, Department of Surgery and Cancer, Division of Cancer, Du Cane Road, London W12 0NN, United KingdombFoundation for Research and Technology Hellas-Institute of Electronic Structure and Laser, N. Plastira 100, 7.
authors.forename: Rosy
authors.initials: R
authors.lastname: Favicchio
authors.affiliation: Foundation for Research and Technology Hellas-Institute of Electronic Structure and Laser, N. Plastira 100, 7100 Heraklion, Crete, Greece.
authors.forename: Stylianos
authors.initials: S
authors.lastname: Psycharakis
authors.affiliation: Central Institute of Mental Health and Heidelberg University, Department of Molecular Biology, Medical Faculty Mannheim, J5, 68159 Mannheim, Germany.
authors.forename: Kai
authors.initials: K
authors.lastname: Schönig
authors.affiliation: Central Institute of Mental Health and Heidelberg University, Department of Molecular Biology, Medical Faculty Mannheim, J5, 68159 Mannheim, Germany.
authors.forename: Dusan
authors.initials: D
authors.lastname: Bartsch
authors.affiliation: Foundation for Research and Technology Hellas-Institute of Molecular Biology and Biotechnology, N. Plastira 100, 7100 Heraklion, Crete, Greece.
authors.forename: Clio
authors.initials: C
authors.lastname: Mamalaki
authors.affiliation: Foundation for Research and Technology Hellas-Institute of Molecular Biology and Biotechnology, N. Plastira 100, 7100 Heraklion, Crete, Greece.
authors.forename: Joseph
authors.initials: J
authors.lastname: Papamatheakis
authors.affiliation: Universidad Carlos III of Madrid, Department of Bioengineering and Aerospace Engineering, 28911 Madrid, SpainfInstituto de Investigación Sanitaria del Hospital Gregorio Marañón, Experimental Medicine and Surgery Unit, 28007 Madrid, Spain.
authors.forename: Jorge
authors.initials: J
authors.lastname: Ripoll
authors.affiliation: Foundation for Research and Technology Hellas-Institute of Electronic Structure and Laser, N. Plastira 100, 7100 Heraklion, Crete, Greece.
authors.forename: Giannis
authors.initials: G
authors.lastname: Zacharakis
mesh_terms: D000818:Animals; D004867:Equipment Design; D005456:Fluorescent Dyes / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D051379:Mice; D008822:Mice, Transgenic; D057054:Molecular Imaging / Q000379:methods*; D061848:Optical Imaging / Q000379:methods*; D015203:Reproducibility of Results; D041622:Tomography, Optical / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes