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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics


  • Editor: Gregory W. Faris
  • Vol. 2, Iss. 3 — Mar. 7, 2007

4Pi microscopy with linear fluorescence excitation

Marion C. Lang, Johann Engelhardt, and Stefan W. Hell  »View Author Affiliations

Optics Letters, Vol. 32, Issue 3, pp. 259-261 (2007)

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Practical 4Pi microscopy has so far exclusively relied on multiphoton excitation of fluorescence, because the nonlinear suppression of contributions from higher-order sidelobes was mandatory for unambiguous axial superresolution. We show that novel lenses of 74 ° semiaperture angle enable biological 4Pi microscopy with regular one-photon fluorescence excitation, thus increasing the signal and reducing system complexity and cost. An axial resolution of 95 nm , corresponding to a more than fourfold improvement over confocal microscopy, is verified in the imaging of microtubules in mammalian cells.

© 2007 Optical Society of America

OCIS Codes
(100.6640) Image processing : Superresolution
(180.1790) Microscopy : Confocal microscopy
(180.2520) Microscopy : Fluorescence microscopy
(180.6900) Microscopy : Three-dimensional microscopy

ToC Category:

Original Manuscript: September 1, 2006
Revised Manuscript: October 21, 2006
Manuscript Accepted: October 26, 2006
Published: January 12, 2007

Virtual Issues
Vol. 2, Iss. 3 Virtual Journal for Biomedical Optics

Marion C. Lang, Johann Engelhardt, and Stefan W. Hell, "4Pi microscopy with linear fluorescence excitation," Opt. Lett. 32, 259-261 (2007)

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