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Journal of Biomedical Optics

Preclinical whole body time domain fluorescence lifetime multiplexing of fluorescent proteins
Author(s): William L. Rice; Anand T. Kumar
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Paper 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.

Paper Details

Date Published: 8 April 2014
PDF: 7 pages
J. Biomed. Opt. 19(4) 046005 doi: 10.1117/1.JBO.19.4.046005
Published in: Journal of Biomedical Optics Volume 19, Issue 4
Show Author Affiliations
William L. Rice, Harvard Medical School (United States)
Anand T. Kumar, Massachusetts General Hospital (United States)


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