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Proceedings Paper

Use of UV-excitation for confocal fluorescence microscopy in a conventional beam-scanning instrument
Author(s): Kjell Carlsson; Karin Mossberg; Johannes P. Helm; Johan Philip
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Paper Abstract

By making only minor modifications, we have adapted a conventional confocal scanning laser microscope for the recording of UV-excited fluorescence. An external argon ion laser provides the wavelengths 334, 351, and 364 nm for specimen illumination. In addition to substituting some optical components to obtain improved transmission and reflection properties in the UV, we have also adjusted the ray-path to compensate for the severe chromatic aberration of most conventional microscope objectives in the UV. We have also tested mirror objectives, which are inherently free from chromatic aberrations. However, since these objectives are not of the immersion-type, and furthermore have rather low numerical apertures, they are of limited value in biomedical applications. Using the modified instrument, we have recorded specimens labeled with AMCA and Fluoro-Gold. At present the instrument is capable of recording optical sections with a thickness of 1.5 micrometers when using an oil-immersion objective with a numerical aperture of 1.25.

Paper Details

Date Published: 26 June 1992
PDF: 16 pages
Proc. SPIE 1660, Biomedical Image Processing and Three-Dimensional Microscopy, (26 June 1992); doi: 10.1117/12.59576
Show Author Affiliations
Kjell Carlsson, Royal Institute of Technology (Sweden)
Karin Mossberg, Royal Institute of Technology (Sweden)
Johannes P. Helm, Royal Institute of Technology (Sweden) and Max Planck Institute for M (Germany)
Johan Philip, Royal Institute of Technology (Sweden)

Published in SPIE Proceedings Vol. 1660:
Biomedical Image Processing and Three-Dimensional Microscopy
Raj S. Acharya; Carol J. Cogswell; Dmitry B. Goldgof, Editor(s)

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