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Three‐dimensional imaging of neurons by confocal fluorescence microscopy
Author(s) -
Carlsson K.,
Wallén P.,
Brodin L.
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.tb04296.x
Subject(s) - optical sectioning , confocal , microscopy , confocal microscopy , optics , light sheet fluorescence microscopy , materials science , resolution (logic) , biological specimen , fluorescence , fluorescence microscope , confocal laser scanning microscopy , microscope , two photon excitation microscopy , scanning confocal electron microscopy , artificial intelligence , computer science , biomedical engineering , physics , medicine
Summary The study of neuronal architecture by means of confocal laser microscopy is described. Optical serial sectioning has been performed on whole‐mount specimens, and the resulting stacks of digitally recorded images have been processed with the help of a computer. Specimen preparation is described, as well as the instrument and its performance. It is shown that the limits in photometric quality are set by photon quantum noise. As both light absorption and scattering was low in the studied specimens, the maximum scanning depth was limited mainly by the working distance of the objectives. Compared with traditional methods, confocal microscopy in combination with digital image processing has the following advantages: (1) a truly three‐dimensional (3‐D) reconstruction is obtained, (2) the specimen remains intact, (3) a higher resolution can be obtained, (4) the process is automated and less time‐consuming and (5) various kinds of data processing are possible.