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Proceedings Paper

4Pi-confocal microscopy of live cells
Author(s): Karsten Bahlmann; Stefan Jakobs; Stefan W. Hell
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Paper Abstract

By coherently adding the spherical wavefronts of two opposing lenses, two-photon excitation 4Pi-confocal fluorescence microscopy has achieved three-dimensional imaging with an axial resolution 3-7 times better than confocal microscopy. So far this improvement was possible only in glycerol-mounted, fixed cells. Here we report 4Pi-confocal microscopy of watery objects and its application to the imaging of live cells. Water immersion 4Pi-confocal microscopy of membrane stained live Escherichia coli bacteria attains a 4.3 fold better axial resolution as compared to the best water immersion confocal microscope. The resolution enhancement results into a vastly improved three-dimensional representation of the bacteria. The first images of live biological samples with an all-directional resolution in the 190-280 nm range are presented here, thus establishing a new resolution benchmark in live cell microscopy.

Paper Details

Date Published: 17 June 2002
PDF: 6 pages
Proc. SPIE 4620, Multiphoton Microscopy in the Biomedical Sciences II, (17 June 2002); doi: 10.1117/12.470687
Show Author Affiliations
Karsten Bahlmann, Massachusetts Institute of Technology (United States)
Stefan Jakobs, Max-Planck-Institute for Biophysical Chemistry (Germany)
Stefan W. Hell, Max-Planck Institute for Biophysical Chemistry (Germany)

Published in SPIE Proceedings Vol. 4620:
Multiphoton Microscopy in the Biomedical Sciences II
Ammasi Periasamy; Peter T. C. So, Editor(s)

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