Abstract

We explore the current limits of the axial resolution of optical sectioning microscopy using a single lens, by combining the resolving power of novel 1.45 numerical aperture oil immersion lenses with superresolving binary aperture filters. We quantify the axial resolution brought about by the increase in semiaperture angle to max á =72.8° and demonstrate an absolute gain in axial resolution through binary pupil filters. Implemented in a confocalized two-photon excitation microscope, the combination of both effectively improves the axial resolution to 330 nm full-width-half-maximum, which is by 34% over that of a standard 1.4 NA system.

© 2002 Optical Society of America

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Appl. Opt.

Appl. Phys. Lett.

C. M. Blanca, J. Bewersdorf and S. W. Hell, �??Single sharp spot in fluorescence microscopy of two opposing lenses, �?? Appl. Phys. Lett. 79, 2321-2323 (2001).
[CrossRef]

Bioimaging

T. Wilson and R. Juskaitis, �??The axial response of confocal microscopes with high numerical aperture objective lenses,�?? Bioimaging 3, 35-38 (1995).
[CrossRef]

J. Biomed. Opt.

N. Huse, A. Schönle and S. W. Hell, �??Z-polarized confocal microscopy,�?? J. Biomed. Opt. 6, 480-484 (2001).
[CrossRef]

J. Opt. Soc. Am. A

Opt. Commun.

S. Quabis, R. Dorn, M. Eberler, O. Glöckl and G. Leuchs, �??Focusing light to a tighter spot,�?? Opt. Commun. 179, 1-7 (2000).
[CrossRef]

M. Martinez-Corral, P. Andres, C. J. Zapata-Rodriguez and M. Kowalczyk, �??Three-dimensional superresolution by annular binary filters, �?? Opt. Commun. 165, 267-278 (1999).
[CrossRef]

Opt. Express

Opt. Lett.

Phys. Rev. Lett.

L. Novotny, M. R. Beversluis, K. S. Youngworth and T. G. Brown, �??Longitudinal Field Modes Probed by Single Molecules,�?? Phys. Rev. Lett. 86, 5251-5254 (2001).
[CrossRef]

Proc. R. Soc. Lond. A

B. Richards and E. Wolf, �??Electromagnetic diffraction in opticel systems II. Structure of the image field in an aplanatic system, �?? Proc. R. Soc. Lond. A 253, 358-379 (1959).

Single Mol.

C. M. Blanca and S. W. Hell, �??Sharp Spherical Focal Spot by Dark Ring 4Pi-Confocal Microscopy, �?? Single Mol. 2, 207-210 (2001).
[CrossRef]

Other

T. Wilson and C. J. R. Sheppard, Theory and Practice of Scanning Optical Microscopy (Academic Press, New York, 1984).

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