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Optical sectioning in confocal fluorescent microscopes
Author(s) -
Wilson T.
Publication year - 1989
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 - 10.1111/j.1365-2818.1989.tb00577.x
Subject(s) - optical sectioning , confocal , fluorescence , microscope , optics , confocal microscopy , fluorescence microscope , microscopy , materials science , wavelength , optical microscope , light sheet fluorescence microscopy , optoelectronics , scanning electron microscope , physics
We discuss the three‐dimensional imaging properties of confocal fluorescence scanning optical microscopes with particular reference to their optical sectioning property. We consider the practically important cases of the use of finite‐sized circular and slit detectors and show how they effect the strength of the sectioning. We find, inter alia, that the performance is always better if the excitation and fluorescent wavelengths are close to each other, indeed as the fluorescent wavelength becomes very large the sectioning disappears altogether. We also compare the strength of the sectioning with that obtained in non‐fluorescent confocal microscopy as this has implications in immuno‐gold labelling studies.