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Journal of Biomedical Optics

Deep tissue fluorescence imaging and in vivo biological applications
Author(s): Viera Crosignani; Alexander S. Dvornikov; Jose S. Aguilar; Chiara Stringari; Robert A. Edwards; William W. Mantulin; Enrico Gratton
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Paper Abstract

We describe a novel technical approach with enhanced fluorescence detection capabilities in two-photon microscopy that achieves deep tissue imaging, while maintaining micron resolution. Compared to conventional two-photon microscopy, greater imaging depth is achieved by more efficient harvesting of fluorescence photons propagating in multiple-scattering media. The system maintains the conventional two-photon microscopy scheme for excitation. However, for fluorescence collection the detection system harvests fluorescence photons directly from a wide area of the turbid sample. The detection scheme relies on a wide area detector, minimal optical components and an emission path bathed in a refractive-index-matching fluid that minimizes emission photon losses. This detection scheme proved to be very efficient, allowing us to obtain high resolution images at depths up to 3 mm. This technique was applied to in vivo imaging of the murine small intestine (SI) and colon. The challenge is to image normal and diseased tissue in the whole live animal, while maintaining high resolution imaging at millimeter depth. In Lgr5-GFP mice, we have been successful in imaging Lgr5-eGFP positive stem cells, present in SI and colon crypt bases.

Paper Details

Date Published: 8 November 2012
PDF: 7 pages
J. Biomed. Opt. 17(11) 116023 doi: 10.1117/1.JBO.17.11.116023
Published in: Journal of Biomedical Optics Volume 17, Issue 11
Show Author Affiliations
Viera Crosignani, Univ. of California, Irvine (United States)
Alexander S. Dvornikov, Univ. of California, Irvine (United States)
Jose S. Aguilar, Univ. of California, Irvine (United States)
Chiara Stringari, Univ. of California, Irvine (United States)
Robert A. Edwards, Univ. of California, Irvine (United States)
William W. Mantulin, Univ. of California, Irvine (United States)
Enrico Gratton, Univ. of California, Irvine (United States)

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