Abstract

Confocal scanning microscopy is widely used for three-dimensional (3-D) visualization of fixed specimens but has found only a limited 3-D reconstruction application for living specimens because the high intensity of the excitation often damages the specimen or causes the fluorescent dye to bleach. Computational optical-sectioning microscopy also suffers from drawbacks because nonconfocal 3-D imaging is fundamentally constrained by an artifactual elongation in the optical axis imposed by the so-called missing cone. We investigate the imaging properties of a new rotating-disk partially confocal scanning microscope (PCSM) that greatly reduces collection time by using multiple apertures for both excitation and detection, effectively working as many confocal microscopes in parallel. We show that this PCSM behaves as a hybrid microscope; near the in-focus plane it behaves near the theoretical optimum for confocal microscopy, and away from this plane its behavior approaches that of a nonconfocal microscope. We also show that the rotating-disk PCSM does not suffer from a missing cone. In fact, the optical transfer function of the theoretically optimal confocal microscope and the rotating-disk PCSM have practically the same bandpass in the spatial-frequency domain.

© 1994 Optical Society of America

Full Article  |  PDF Article
OSA Recommended Articles
Calculation of three-dimensional optical transfer function for a confocal scanning fluorescent microscope

Shigeharu Kimura and Chusuke Munakata
J. Opt. Soc. Am. A 6(7) 1015-1019 (1989)

References

You do not have subscription access to this journal. Citation lists with outbound citation links are available to subscribers only. You may subscribe either as an OSA member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access OSA Member Subscription

Cited By

You do not have subscription access to this journal. Cited by links are available to subscribers only. You may subscribe either as an OSA member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access OSA Member Subscription

Figures (9)

You do not have subscription access to this journal. Figure files are available to subscribers only. You may subscribe either as an OSA member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access OSA Member Subscription

Equations (33)

You do not have subscription access to this journal. Equations are available to subscribers only. You may subscribe either as an OSA member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access OSA Member Subscription

Metrics

You do not have subscription access to this journal. Article level metrics are available to subscribers only. You may subscribe either as an OSA member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access OSA Member Subscription