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- W2093597740 abstract "Over the past two decades, multiphoton microscopy (MPM) has launched a revolution in high-resolution in vivo imaging with its combination of molecular contrast, optical sectioning and depth penetration. However, as with other optical microscopies, the imaging depth of MPM is ultimately limited by the scattering of both excitation light and emitted fluorescence. Since multiphoton excitation is largely confined to a diffraction-limited focal volume, MPM can theoretically utilize any fluorescence as signal and most collection schemes seek to collect as many photons as possible. The collected fraction depends on the choice of objective lens and the design of the post-objective light collection path, as well as the optical properties of the specimen. In particular, this fraction decreases significantly in turbid samples. Recently, there has been a concerted effort by lens manufacturers to develop objective lenses that are specially tailored for MPM and aim to collect as much scattered light as possible. In order to assess the advancements made by this “second generation” of MP objectives, we directly measure the collected fraction from a sample consisting of a fluorescein layer below a solution of scattering microspheres. Trans- and epi-fluorescence are collected in separate channels to give a relative measure of the collected fraction and the dependence on sample scattering is compared for five different lenses. We also observe that the angular divergence of the light cone exiting the objective back increases with sample scattering- a practical detail that has implications on design of collection pathways." @default.
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- W2093597740 date "2010-01-01" @default.
- W2093597740 modified "2023-10-16" @default.
- W2093597740 title "A Comparison of Objective Lenses for Multiphoton Microscopy: Improved Epifluorescence Collection from Turbid Samples" @default.
- W2093597740 doi "https://doi.org/10.1016/j.bpj.2009.12.951" @default.
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