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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 15, Iss. 5 — Mar. 5, 2007
  • pp: 2459–2467

4Pi microscopy of type A with 1-photon excitation in biological fluorescence imaging

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

Optics Express, Vol. 15, Issue 5, pp. 2459-2467 (2007)

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We demonstrate that oil immersion lenses with a semi-aperture angle ≥ 74° enable 4Pi confocal fluorescence microscopy of type A with 1-photon excitation. The axial sidelobes amount to < 50 % of the main diffraction maximum, implying that lobe induced artifacts can be removed from the image data. The advancement reported herein enables a relative inexpensive implementation of 4Pi microscopy, providing axially superresolved 3D-imaging in transparent samples. As an example, we show dual-color 4Pi images of double stained Golgi stacks in a mammalian cell with 110 nm axial resolution. The resolution can be further enhanced to values slightly below 100 nm by image deconvolution.

© 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: January 10, 2007
Manuscript Accepted: February 19, 2007
Published: March 5, 2007

Virtual Issues
Vol. 2, Iss. 4 Virtual Journal for Biomedical Optics

Marion Lang, Tobias Müller, Johann Engelhardt, and Stefan W. Hell, "4Pi microscopy of type A with 1-photon excitation in biological fluorescence imaging," Opt. Express 15, 2459-2467 (2007)

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