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Phys. Rev. Lett. 88, 163901 (2002)

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Focal Spots of Size /23 Open Up Far-Field Florescence Microscopy at 33 nm Axial Resolution

Marcus Dyba and Stefan W. Hell

High Resolution Optical Microscopy Group, Max-Planck-Institute for Biophysical Chemistry, 37070 Göttingen, Germany

(Received 19 September 2001; published 4 April 2002)

We report spots of excited molecules of 33 nm width created with focused light of = 760  nm wavelength and conventional optics along the optic axis. This is accomplished by exciting the molecules with a femtosecond pulse and subsequent depletion of their excited state with red-shifted, picosecond-pulsed, counterpropagating, coherent light fields. The /23 ratio constitutes what is to our knowledge the sharpest spatial definition attained with freely propagating electromagnetic radiation. The sub-diffraction spots enable for the first time far-field fluorescence microscopy with resolution at the tens of nanometer scale, as demonstrated in images of membranes of bacillus megaterium. ©2002 The American Physical Society

URL: http://link.aps.org/abstract/PRL/v88/e163901
doi:10.1103/PhysRevLett.88.163901
PACS: 42.30.-d, 41.90.+e, 42.25.-p, 42.79.-e        Additional Information

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