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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 16, Iss. 26 — Dec. 22, 2008
  • pp: 21843–21848

Real-time extended depth of field microscopy

Edward J. Botcherby, Martin J. Booth, Rimas Juškaitis, and Tony Wilson  »View Author Affiliations


Optics Express, Vol. 16, Issue 26, pp. 21843-21848 (2008)
http://dx.doi.org/10.1364/OE.16.021843


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Abstract

We describe an optical microscope system whose focal setting can be changed quickly without moving the objective lens or specimen. Using this system, diffraction limited images can be acquired from a wide range of focal settings without introducing optical aberrations that degrade image quality. We combine this system with a real time Nipkow disc based confocal microscope so as to permit the acquisition of extended depth of field images directly in a single frame of the CCD camera. We also demonstrate a simple modification that enables extended depth of field images to be acquired from different angles of perspective, where the angle can be changed over a continuous range by the user in real-time.

© 2008 Optical Society of America

OCIS Codes
(180.6900) Microscopy : Three-dimensional microscopy

ToC Category:
Microscopy

History
Original Manuscript: September 8, 2008
Revised Manuscript: November 20, 2008
Manuscript Accepted: November 24, 2008
Published: December 17, 2008

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

Citation
Edward J. Botcherby, Martin J. Booth, Rimas Juskaitis, and Tony Wilson, "Real-time extended depth of field microscopy," Opt. Express 16, 21843-21848 (2008)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-16-26-21843


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References

  1. T. Wilson and C. J. R. Sheppard, Theory and Practice of Scanning Optical Microscopy (Academic Press, London, 1984).
  2. M. A. A. Neil, R. Ju¡skaitis, T. Wilson, Z. J. Laczik, and V. Sarafis, "Optimized pupil-plane filters for confocal microscope point-spread function engineering," Opt. Lett. 25,245-247 (2000). [CrossRef]
  3. E. Botcherby, R. Ju¡skaitis, and T. Wilson, "Scanning two photon fluorescence microscopy with extended depth of field," Opt. Comm. 268,253-260 (2006). [CrossRef]
  4. N. George and W. Chi, "Extended depth of field using a logarithmic asphere," J. Opt. A: Pure Appl. Opt. 5,S157-S163 (2003). [CrossRef]
  5. E. R. Dowski and W. T. Cathey, "Extended depth of field through wave-front coding," Appl. Opt. 34,1859-1866 (1995). [CrossRef] [PubMed]
  6. M. Born and E. Wolf, Principles of Optics (Pergamon Press, 6th edition, 1983).
  7. E. Botcherby, R. Ju¡skaitis, M. Booth, and T. Wilson, "Aberration-free optical refocusing in high numerical aperture microscopy," Opt. Lett. 32,2007-2009 (2007). [CrossRef] [PubMed]
  8. E. Botcherby, R. Ju¡skaitis, M. Booth, and T. Wilson, "An optical technique for remote focusing in microscopy," Opt. Comm. 281,880-887 (2008). [CrossRef]

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