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Research Papers: Imaging

Lifetime-based tomographic multiplexing

[+] Author Affiliations
Scott B. Raymond

The Harvard-MIT Division of Health Sciences and Technology, 114 16th Street, Charlestown, Massachusetts 02129

David A. Boas

Massachusetts General Hospital, Department of Radiology, Athinoula A. Martinos Center for Biomedical Imaging, 149 13th Street, Charlestown, Massachusetts 02129

Brian J. Bacskai

Massachusetts General Hospital, Department of Neurology, Alzheimer’s Research Unit, 114 16th Street, Charlestown, Massachusetts 02129

Anand T. N. Kumar

Massachusetts General Hospital, Department of Radiology, Athinoula A. Martinos Center for Biomedical Imaging, 149 13th Street, Charlestown, Massachusetts 02129

J. Biomed. Opt. 15(4), 046011 (August 06, 2010). doi:10.1117/1.3469797
History: Received March 21, 2010; Revised May 16, 2010; Accepted June 01, 2010; Published August 06, 2010; Online August 06, 2010
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Near-infrared (NIR) fluorescence tomography of multiple fluorophores has previously been limited by the bandwidth of the NIR spectral regime and the broad emission spectra of most NIR fluorophores. We describe in vivo tomography of three spectrally overlapping fluorophores using fluorescence lifetime-based separation. Time-domain images are acquired using a voltage-gated, intensified charge-coupled device (CCD) in free-space transmission geometry with 750nm Ti:sapphire laser excitation. Lifetime components are fit from the asymptotic portion of fluorescence decay curve and reconstructed separately with a lifetime-adjusted forward model. We use this system to test the in vivo lifetime multiplexing suitability of commercially available fluorophores, and demonstrate lifetime multiplexing in solution mixtures and in nude mice. All of the fluorophores tested exhibit nearly monoexponential decays, with narrow in vivo lifetime distributions suitable for lifetime multiplexing. Quantitative separation of two fluorophores with lifetimes of 1.1 and 1.37ns is demonstrated for relative concentrations of 1:5. Finally, we demonstrate tomographic imaging of two and three fluorophores in nude mice with fluorophores that localize to distinct organ systems. This technique should be widely applicable to imaging multiple NIR fluorophores in 3-D.

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© 2010 Society of Photo-Optical Instrumentation Engineers

Citation

Scott B. Raymond ; David A. Boas ; Brian J. Bacskai and Anand T. N. Kumar
"Lifetime-based tomographic multiplexing", J. Biomed. Opt. 15(4), 046011 (August 06, 2010). ; http://dx.doi.org/10.1117/1.3469797


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