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Journal of Biomedical Optics

Emission spectra of bioluminescent reporters and interaction with mammalian tissue determine the sensitivity of detection in vivo
Author(s): Hui Zhao; Timothy C. Doyle; Olivier Coquoz; Flora Kalish; Bradley W. Rice; Christopher H. Contag
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Paper Abstract

In vivo bioluminescence imaging depends on light emitted by luciferases in the body overcoming the effect of tissue attenuation. Understanding this relationship is essential for detection and quantification of signal. We have studied four codon optimized luciferases with different emission spectra, including enzymes from firefly (FLuc), click beetle (CBGr68, CBRed) and Renilla reniformins (hRLuc). At 25°C, the in vitro λmax of these reporters are 578, 543, 615, and 480 nm, respectively; at body temperature, 37°C, the brightness increases and the firefly enzyme demonstrates a 34-nm spectral red shift. Spectral shifts and attenuation due to tissue effects were evaluated using a series of 20-nm bandpass filters and a cooled charge-coupled device (CCD) camera. Attenuation increased and the spectra of emitted light was red shifted for signals originating from deeper within the body relative to superficial origins. The tissue attenuation of signals from CBGr68 and hRLuc was greater than from those of Fluc and CBRed. To further probe tissue effects, broad spectral emitters were created through gene fusions between CBGr68 and CBRed. These resulted in enzymes with broader emission spectra, featuring two peaks whose intensities are differentially affected by temperature and tissue depth. These spectral measurement data allow for improved understanding of how these reporters can be

Paper Details

Date Published: 1 July 2005
PDF: 9 pages
J. Biomed. Opt. 10(4) 041210 doi: 10.1117/1.2032388
Published in: Journal of Biomedical Optics Volume 10, Issue 4
Show Author Affiliations
Hui Zhao, Stanford Linear Accelerator Center (United States)
Timothy C. Doyle, Stanford Linear Accelerator Center (United States)
Olivier Coquoz, Xenogen Corp. (United States)
Flora Kalish, Stanford Linear Accelerator Center (United States)
Bradley W. Rice, Xenogen Corp. (United States)
Christopher H. Contag, Stanford Linear Accelerator Center (United States)

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