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Journal of Biomedical Optics

Performance of the red-shifted fluorescent proteins in deep-tissue molecular imaging applications
Author(s): Nikolaos Deliolanis; Randa Kasmieh; Thomas Wurdinger; Bakhos A. Tannous; Khalid Shah; Vasilis Ntziachristos
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Paper Abstract

The discovery of new fluorescent proteins (FPs) that emit in the far-red part of the spectrum, where light absorption from tissue is significantly lower than in the visible, offers the possibility for noninvasive biological interrogation at the entire organ or small animal level in vivo. The performance of FPs in deep-tissue imaging depends not only on their optical characteristics, but also on the wavelength-dependent tissue absorption and the depth of the fluorescence activity. To determine the optimal choice of FP and illumination wavelength, we compared the performance of five of the most promising FPs: tdTomato, mCherry, mRaspberry, mPlum, and Katushka. We experimentally measured the signal strength through mice and employed theoretical predictions to obtain an understanding of the performance of different illumination scenarios, especially as they pertain to tomographic imaging. It was found that the appropriate combination of red-shifted proteins and illumination wavelengths can improve detection sensitivity in small animals by at least two orders of magnitude compared with green FP. It is also shown that the steep attenuation change of the hemoglobin spectrum around the 600-nm range may significantly affect the detection sensitivity and, therefore, necessitates the careful selection of illumination wavelengths for optimal imaging performance.

Paper Details

Date Published: 1 July 2008
PDF: 9 pages
J. Biomed. Opt. 13(4) 044008 doi: 10.1117/1.2967184
Published in: Journal of Biomedical Optics Volume 13, Issue 4
Show Author Affiliations
Nikolaos Deliolanis, Massachusetts General Hospital (United States)
Randa Kasmieh, Massachusetts General Hospital (United States)
Thomas Wurdinger, Massachusetts General Hospital (United States)
Bakhos A. Tannous, Massachusetts General Hospital (United States)
Khalid Shah, Massachusetts General Hospital (United States)
Vasilis Ntziachristos, Massachusetts General Hospital (United States)

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