Research Papers: Imaging

Preclinical whole body time domain fluorescence lifetime multiplexing of fluorescent proteins

[+] Author Affiliations
William L. Rice

Massachusetts General Hospital, Harvard Medical School, Athinoula A. Martinos Center for Biomedical Imaging, 149 13 Street, Charlestown, Massachusetts 02129

Anand T. N. Kumar

Massachusetts General Hospital, Harvard Medical School, Athinoula A. Martinos Center for Biomedical Imaging, 149 13 Street, Charlestown, Massachusetts 02129

J. Biomed. Opt. 19(4), 046005 (Apr 08, 2014). doi:10.1117/1.JBO.19.4.046005
History: Received February 6, 2014; Revised March 12, 2014; Accepted March 17, 2014
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Abstract.  The application of time domain (TD) fluorescence lifetime multiplexing for the detection of fluorescent proteins (FPs) in whole animals, in the presence of a strong background tissue autofluorescence and excitation light leakage is discussed. Tissue autofluorescence (AF) exhibits a nonexponential temporal response, distinct from the mono-exponential decay of FPs. This allows a direct separation of FP fluorescence from AF using a dual basis function approach. We establish the detection limits of this approach using in vitro and in vivo measurements. We also demonstrate, using an experimental model of lymph node metastasis, that FP-AF lifetime multiplexing provides a greater than 30-fold improvement in contrast-to-background ratio compared with continuous wave data. In addition, we show that TD detection can simultaneously discriminate between up to three red shifted FPs placed under the skin of a nude mouse based on their distinct fluorescence lifetimes.

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© 2014 Society of Photo-Optical Instrumentation Engineers

Citation

William L. Rice and Anand T. N. Kumar
"Preclinical whole body time domain fluorescence lifetime multiplexing of fluorescent proteins", J. Biomed. Opt. 19(4), 046005 (Apr 08, 2014). ; http://dx.doi.org/10.1117/1.JBO.19.4.046005


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