8 November 2012 Deep tissue fluorescence imaging and in vivo biological applications
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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.
© 2012 Society of Photo-Optical Instrumentation Engineers (SPIE)
Viera Crosignani, Viera Crosignani, Alexander S. Dvornikov, Alexander S. Dvornikov, Jose S. Aguilar, Jose S. Aguilar, Chiara Stringari, Chiara Stringari, Robert A. Edwards, Robert A. Edwards, William W. Mantulin, William W. Mantulin, Enrico Gratton, Enrico Gratton, } "Deep tissue fluorescence imaging and in vivo biological applications," Journal of Biomedical Optics 17(11), 116023 (8 November 2012). https://doi.org/10.1117/1.JBO.17.11.116023 . Submission:

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