Preclinical whole body time domain fluorescence lifetime multiplexing of fluorescent proteins

William L. Rice; Anand T. Kumar

doi:10.1117/1.JBO.19.4.046005

8 April 2014 Preclinical whole body time domain fluorescence lifetime multiplexing of fluorescent proteins

William L. Rice, Anand T. Kumar

Author Affiliations +

Journal of Biomedical Optics, Vol. 19, Issue 4, 046005 (April 2014). https://doi.org/10.1117/1.JBO.19.4.046005

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.

Citation Download Citation

William L. Rice and Anand T. Kumar "Preclinical whole body time domain fluorescence lifetime multiplexing of fluorescent proteins," Journal of Biomedical Optics 19(4), 046005 (8 April 2014). https://doi.org/10.1117/1.JBO.19.4.046005

Published: 8 April 2014

ACCESS THE FULL ARTICLE

JOURNAL ARTICLE
7 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY

GET CITATION

CITATIONS

Cited by 10 scholarly publications.

Explore citations on Lens.org

RIGHTS & PERMISSIONS

Get copyright permission Get copyright permission on Copyright Marketplace

KEYWORDS

Atrial fibrillation

Luminescence

Green fluorescent protein

Continuous wave operation

Multiplexing

Skin

Tissues

Show All Keywords

RELATED CONTENT

Simultaneous detection of tissue autofluorescence decay distribution and time gated...
Proceedings of SPIE (May 06 2015)

Spectral unmixing of multi color tissue specific in vivo fluorescence...
Proceedings of SPIE (July 11 2007)

Use of GFP for in vivo imaging concepts and...
Proceedings of SPIE (February 13 2008)

Revisiting optical clearing with dimethyl sulfoxide (DMSO) in vitro...
Proceedings of SPIE (February 24 2009)

Clinical applications of in vivo fluorescence confocal laser scanning microscopy
Proceedings of SPIE (March 05 2008)

In vivo time-resolved autofluorescence measurements on human skin
Proceedings of SPIE (February 22 2006)

Subscribe to Digital Library

Receive Erratum Email Alert

Show All Keywords

Keywords/Phrases

Search In:

Publication Years