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- W2022146375 abstract "We evaluate the point spread function (PSF) of the two-color excitation (2CE) microscope when the confocality condition is not strictly satisfied by the two focused excitation beams which are separated by an angle θ≠0. The 2CE wavelengths λ1 and λ2 are related to the single-photon excitation wavelength λe of the sample according to: 1/λe=1/λ1+1/λ2. Spatio-temporal simultaneity for the two excitation beams is essential in 2CE (λ1≠λ2). Only aberration-free misalignment of the geometrical foci is considered and the degradation of the peak 2CE fluorescence intensity (F2cp) is determined as a function of the focus separation for θ=0,π/2 and π. The deterioration of the fluorescence PSF is also evaluated using the Linfoot's criteria of fidelity, structural content and correlation quality. We compare the 2CE PSF behavior with that of the two-photon excitation (2PE) fluorescence (λ1=λ2) under the same misalignment settings. At θ=π/2, the F2cp degrades less rapidly than the peak 2PE fluorescence intensity (F2pp) for misalignments that are confined within the focal plane of the focusing lens L1 for λ1. At θ=π,F2cp and F2pp degrade in a similar manner. For misalignments along the optical axis of L1 and at θ=π/2, the F2cp degrades monotonically while the F2pp degradation is accompanied by fluctuations caused by two-beam interference. Our calculations reveal that 2CE imaging is more resilient to the ill-effects of misalignment than the two-beam 2PE." @default.
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- W2022146375 date "2002-06-01" @default.
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- W2022146375 title "Confocality condition in two-color excitation microscopy with two focused excitation beams" @default.
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- W2022146375 doi "https://doi.org/10.1016/s0030-4018(02)01530-4" @default.
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