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Deliolanis, N.C.* ; Kasmieh, R.* ; Wurdinger, T.* ; Tannous, B.A.* ; Shah, K.* ; Ntziachristos, V.*

Performance of the red-shifted fluorescent proteins in deep-tissue molecular imaging applications.

J. Biomed. Opt. 13, 0440081-0440089 (2008)
DOI
Open Access Gold as soon as Publ. Version/Full Text is submitted to ZB.
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.
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Publication type Article: Journal article
Document type Scientific Article
Keywords fluorescent proteins; optical tomography; whole-body imaging
ISSN (print) / ISBN 1083-3668
e-ISSN 1560-2281
Journal Journal of Biomedical Optics
Quellenangaben Volume: 13, Issue: 4, Pages: 0440081-0440089 Article Number: , Supplement: ,
Publisher SPIE
Publishing Place Bellingham, WA
Reviewing status Peer reviewed
Institute(s) Institute of Biological and Medical Imaging (IBMI)