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Proceedings Paper

Compact 4pi beam scanning microscope for imaging cells with a sevenfold increase in axial resolution
Author(s): Hilmar F. Gugel; Reiner Rygiel; Rafael Storz
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Paper Abstract

Confocal microscopy is the method of choice in biological 3D-imaging, however, the axial resolution is limited to ~500 nm. During the last decade it has been successfully demonstrated that the axial resolution can be substantially improved with 4Pi microscopy. We report a 4Pi microscope realized as a fast beam scanning system consisting of a 4Pi-module linked to a state-of-the-art confocal microscope. As a result, the advantages of the confocal system such as scanning, sensitive multicolor detection and imaging speed are combined with the superior resolution of 4Pi microscopy. This novel microscope is eminently suited for biological applications. It is designed both for single-photon and for two-photon picosecond excitation, and also enables joint coherent illumination and detection, i.e. 4Pi type C. The superior PSF and OTF of the system enable 80 nm axial resolution in cells mounted in aqueous media. We present the optical design of the system and demonstrate an up to 7-fold improved optical sectioning in live cells.

Paper Details

Date Published: 30 March 2005
PDF: 6 pages
Proc. SPIE 5700, Multiphoton Microscopy in the Biomedical Sciences V, (30 March 2005); doi: 10.1117/12.618525
Show Author Affiliations
Hilmar F. Gugel, Leica Microsystems CMS GmbH (Germany)
Reiner Rygiel, Leica Microsystems CMS GmbH (Germany)
Rafael Storz, Leica Microsystems CMS GmbH (Germany)

Published in SPIE Proceedings Vol. 5700:
Multiphoton Microscopy in the Biomedical Sciences V
Ammasi Periasamy; Peter T. C. So, Editor(s)

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