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  • Bahlmann, K.,Jakobs, S. and Hell, S.W.: 4Pi-confocal microscopy of live cells

Bibliographic Data

  • Authors: Bahlmann, K.,Jakobs, S. and Hell, S.W.
  • Title: 4Pi-confocal microscopy of live cells
  • Journal: Ultramicroscopy
  • Volume: 87
  • Issue: 3
  • Volume: 155-164
  • DOI: 10.1016/s0304-3991(00)00092-9

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 of 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. (C) 2001 Elsevier Science B.V. All rights reserved.