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Image enhancement in ultramicroscopy by improved laser light sheets
Author(s) -
Saghafi Saiedeh,
Becker Klaus,
Jährling Nina,
Richter Melanie,
Kramer Edgar R.,
Dodt HansUlrich
Publication year - 2010
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.201000047
Subject(s) - optics , numerical aperture , lens (geology) , aperture (computer memory) , laser , materials science , rayleigh length , light sheet fluorescence microscopy , focal point , optical sectioning , resolution (logic) , beam (structure) , microscopy , laser beams , physics , cardinal point , scanning confocal electron microscopy , computer science , wavelength , artificial intelligence , acoustics
In the majority of implementations of light sheet microscopy, such as ultramicroscopy, the laser beam illuminating the specimen is truncated by a slit aperture before it is focused to a light sheet by a single cylindrical lens. A light sheet generated in this way can be made very thin near to the focal point, but unfortunately its Rayleigh range is severely limited. This problem can be partially solved by using a smaller slit aperture. However, this also causes a major loss in power, a severe broadening of the beam waist, and thus a significant loss of resolution along the detection axis. We developed improved light‐sheet‐generation optics, which provide longer Raleigh ranges, whilst retaining beam waists comparable to our standard system with one cylindrical lens. Using the modified system we achieved a marked improvement in the resolution of ultramicroscopy reconstructions of representative biological specimens. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)