期刊
OPTICS EXPRESS
卷 19, 期 16, 页码 15348-15362出版社
OPTICAL SOC AMER
DOI: 10.1364/OE.19.015348
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- National Center for Research Resources (NCRR) [TL 1 RR024137]
- National Insitute of Biomedical Imaging and Bioengineering (NIBIB) [R21EB00770]
Understanding fluorescence propagation through a multiphoton microscope is of critical importance in designing high performance systems capable of deep tissue imaging. Optical models of a scattering tissue sample and the Olympus 20X 0.95NA microscope objective were used to simulate fluorescence propagation as a function of imaging depth for physiologically relevant scattering parameters. The spatio-angular distribution of fluorescence at the objective back aperture derived from these simulations was used to design a simple, maximally efficient post-objective fluorescence collection system. Monte Carlo simulations corroborated by data from experimental tissue phantoms demonstrate collection efficiency improvements of 50% - 90% over conventional, non-optimized fluorescence collection geometries at large imaging depths. Imaging performance was verified by imaging layer V neurons in mouse cortex to a depth of 850 mu m. (C) 2011 Optical Society of America
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