4Pi-confocal microscopy of live cells

Karsten Bahlmann; Stefan Jakobs; Stefan W. Hell

doi:10.1117/12.470687

17 June 2002 4Pi-confocal microscopy of live cells

Karsten Bahlmann, Stefan Jakobs, Stefan W. Hell

Proceedings Volume 4620, Multiphoton Microscopy in the Biomedical Sciences II; (2002) https://doi.org/10.1117/12.470687
Event: International Symposium on Biomedical Optics, 2002, San Jose, CA, United States

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.

Citation Download Citation

Karsten Bahlmann, Stefan Jakobs, and Stefan W. Hell "4Pi-confocal microscopy of live cells", Proc. SPIE 4620, Multiphoton Microscopy in the Biomedical Sciences II, (17 June 2002); https://doi.org/10.1117/12.470687

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