Pi - confocal microscopy of live cells
نویسندگان
چکیده
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 threedimensional 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. # 2001 Elsevier Science B.V. All rights reserved. PACS: 87.64; 42.30.d; 42.62.Be; 42.79. e
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