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Comparison of the axial resolution of practical Nipkow‐disk confocal fluorescence microscopy with that of multifocal multiphoton microscopy: theory and experiment
Author(s) -
Egner A.,
Andresen V.,
Hell S. W.
Publication year - 2002
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1046/j.1365-2818.2002.01001.x
Subject(s) - optical sectioning , confocal , microscope , confocal microscopy , microscopy , optics , two photon excitation microscopy , excitation wavelength , resolution (logic) , excitation , fluorescence microscope , materials science , super resolution microscopy , fluorescence , fluorescence lifetime imaging microscopy , wavelength , scanning confocal electron microscopy , physics , computer science , artificial intelligence , quantum mechanics
Summary We compare the axial sectioning capability of multifocal confocal and multifocal multiphoton microscopy in theory and in experiment, with particular emphasis on the background arising from the cross‐talk between adjacent imaging channels. We demonstrate that a time‐multiplexed non‐linear excitation microscope exhibits significantly less background and therefore a superior axial resolution as compared to a multifocal single‐photon confocal system. The background becomes irrelevant for thin (< 15 µm) and sparse fluorescent samples, in which case the confocal parallelized system exhibits similar or slightly better sectioning behaviour due to its shorter excitation wavelength. Theoretical and experimental axial responses of practically implemented microscopes are given.