21 February 2017 Absorption characterization of immersion medium for multiphoton microscopy at the 1700nm window
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Abstract
Larger imaging depth is the quest of almost all the imaging modalities, including multiphoton microscopy (MPM). Recently, it has been domonstrated that excitation at the 1700-nm helps extending imaging depth in MPM, optical coherence tomography, as well as photoacoustic imaging compared with excitation at other wavelengths. In MPM, immersion objective lenses with high numerical aperture (NA) are typically used to achieve better signal resolution, higer signal collection efficiency, and stronger signal generation. Although physically short (~mm), this extra optical path length traversed by the excitation light inevitably introduces absorption of the excitation light, and as a result leads to a decrease in the signal generation. Here we demonstrate experimental characterization of absorption spectrum of various immersion media at the 1700-nm window, including water (H2O), deuterium oxide (D2O), and several brands of immersion oil. Our results identify either the best immersion medium for a specific wavelength, or the best wavelength for a specific immersion medium at the 1700-nm window. Furthermore, through quantitative MPM experiments comparing different immersion media, we show that the MPM signal levels can be enhanced by more than ten fold simply by selecting the proper immersion medium, in good agreement with theoretical expectation based on the absorption measurement. Our results will offer guidelines for signal optimization in MPM at the 1700-nm window.
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Wenhui Wen, Ping Qiu, "Absorption characterization of immersion medium for multiphoton microscopy at the 1700nm window", Proc. SPIE 10069, Multiphoton Microscopy in the Biomedical Sciences XVII, 1006929 (21 February 2017); doi: 10.1117/12.2248528; https://doi.org/10.1117/12.2248528
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