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Applied Optics

Applied Optics

APPLICATIONS-CENTERED RESEARCH IN OPTICS

  • Vol. 39, Iss. 10 — Apr. 1, 2000
  • pp: 1652–1658

Polarization effects in 4Pi confocal microscopy studied with water-immersion lenses

Karsten Bahlmann and Stefan W. Hell  »View Author Affiliations


Applied Optics, Vol. 39, Issue 10, pp. 1652-1658 (2000)
http://dx.doi.org/10.1364/AO.39.001652


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Abstract

We studied the effect of electric field orientation on the point-spread function (PSF) of a 4Pi microscope. We show that in a standard 4Pi arrangement the orientation of the field can be used for changing between constructive- and destructive-mode 4Pi microscopy. The effect is counteracted by introduction of a phase shift of π into one of the half-arms. This compensation is compulsory during illumination with unpolarized or circularly polarized light. By performing our experiments with 1.2-N.A. water-immersion lenses, we demonstrate that water immersion is suitable for 4Pi confocal microscopy. At a two-photon excitation wavelength of 1064 nm, the water 4Pi confocal PSF features an axial lobe of 40% above and below the focal plane, which, by linear filtering, can be unambiguously removed. The measured axial full width at half-maximum of the PSF is 240 nm. This is 4.3 times narrower than its single-lens confocal counterpart. The 4Pi confocal microscope sets a new resolution benchmark in three-dimensional imaging of watery samples.

© 2000 Optical Society of America

OCIS Codes
(110.0180) Imaging systems : Microscopy
(110.6880) Imaging systems : Three-dimensional image acquisition
(170.1790) Medical optics and biotechnology : Confocal microscopy
(170.6900) Medical optics and biotechnology : Three-dimensional microscopy
(190.4180) Nonlinear optics : Multiphoton processes

Citation
Karsten Bahlmann and Stefan W. Hell, "Polarization effects in 4Pi confocal microscopy studied with water-immersion lenses," Appl. Opt. 39, 1652-1658 (2000)
http://www.opticsinfobase.org/ao/abstract.cfm?URI=ao-39-10-1652

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