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Research Papers: Imaging

Deep tissue fluorescence imaging and in vivo biological applications

[+] Author Affiliations
Viera Crosignani, Alexander Dvornikov, Jose S Aguilar, Chiara Stringari

University of California at Irvine, Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, Irvine, California 92697

Robert Edwards

University of California at Irvine, School of Medicine, Department of Pathology and Laboratory Medicine, Irvine, California 92697

William W. Mantulin, Enrico Gratton

University of California at Irvine, Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, Irvine, California 92697

University of California at Irvine, Beckman Laser Institute, Irvine, California 92697

J. Biomed. Opt. 17(11), 116023 (Nov 08, 2012). doi:10.1117/1.JBO.17.11.116023
History: Received May 17, 2012; Revised August 28, 2012; Accepted October 16, 2012
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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.

Figures in this Article
© 2012 Society of Photo-Optical Instrumentation Engineers

Citation

Viera Crosignani ; Alexander Dvornikov ; Jose S Aguilar ; Chiara Stringari ; Robert Edwards, et al.
"Deep tissue fluorescence imaging and in vivo biological applications", J. Biomed. Opt. 17(11), 116023 (Nov 08, 2012). ; http://dx.doi.org/10.1117/1.JBO.17.11.116023


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