1. Epifluorescence collection in two-photon microscopy
- Author
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Jerome Mertz, Emmanuel Beaurepaire, Laboratoire de Neurophysiologie et Nouvelles Microscopies (U1128), Université Paris Descartes - Paris 5 (UPD5) - Institut National de la Santé et de la Recherche Médicale (INSERM), and Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)
- Subjects
[PHYS.PHYS.PHYS-OPTICS] Physics [physics]/Physics [physics]/Optics [physics.optics] ,MESH: Photons ,Optics and Photonics ,Photon ,[SDV.IB.IMA]Life Sciences [q-bio]/Bioengineering/Imaging ,Materials Science (miscellaneous) ,Monte Carlo method ,Physics::Optics ,MESH: Microscopy, Fluorescence ,Field of view ,MESH: Monte Carlo Method ,01 natural sciences ,Industrial and Manufacturing Engineering ,Diffusion ,010309 optics ,03 medical and health sciences ,Optics ,MESH: Computer Simulation ,Two-photon excitation microscopy ,0103 physical sciences ,Scattering, Radiation ,Computer Simulation ,MESH: Scattering, Radiation ,Business and International Management ,MESH: Models, Theoretical ,Photon diffusion ,030304 developmental biology ,Physics ,[PHYS.PHYS.PHYS-OPTICS]Physics [physics]/Physics [physics]/Optics [physics.optics] ,Photons ,0303 health sciences ,Geometrical optics ,business.industry ,MESH: Diffusion ,Models, Theoretical ,Numerical aperture ,[SDV.IB.IMA] Life Sciences [q-bio]/Bioengineering/Imaging ,Microscopy, Fluorescence ,Light sheet fluorescence microscopy ,business ,MESH: Optics and Photonics ,Monte Carlo Method - Abstract
International audience; We present a simple model to describe epifluorescence collection in two-photon microscopy when one images in a turbid slab with an objective. Bulk and surface scattering determine the spatial and angular distributions of the outgoing fluorescence photons at the slab surface, and geometrical optics determines how efficiently the photons are collected. The collection optics are parameterized by the objective's numerical aperture and working distance and by an effective collection field of view. We identify the roles of each of these parameters and provide simple rules of thumb for the optimization of the epifluorescence collection efficiency. Analytical results are corroborated by Monte Carlo simulation.
- Published
- 2002
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