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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics


  • Editors: Andrew Dunn and Anthony Durkin
  • Vol. 7, Iss. 6 — May. 25, 2012

Characterizing the 3-D field distortions in low numerical aperture fluorescence zooming microscope

Praveen Pankajakshan, Zvi Kam, Alain Dieterlen, and Jean-Christophe Olivo-Marin  »View Author Affiliations

Optics Express, Vol. 20, Issue 9, pp. 9876-9889 (2012)

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In this article, we characterize the lateral field distortions in a low numerical aperture and large field-of-view (FOV) fluorescence imaging system. To this end, we study a commercial fluorescence MACROscope setup, which is a zooming microscope. The versatility of this system lies in its ability to image at different zoom ranges, so that sample preparations can be examined in three-dimensions, at cellular, organ and whole body levels. Yet, we found that the imaging system’s optics are optimized only for high magnifications where the observed FOV is small. When we studied the point-spread function (PSF) by using fluorescent polystyrene beads as “guide-stars”, we noticed that the PSF is spatially varying due to field distortions. This variation was found to be laterally symmetrical and the distortions were found to increase with the distance from the center of the FOV. In this communication, we investigate the idea of using the field at the back focal plane of an optical system for characterizing distortions. As this field is unknown, we develop a theoretical framework to retrieve the amplitude and phase of the field at the back focal pupil plane, from the empirical bead images. By using the retrieved amplitude, we can understand and characterize the underlying cause of these distortions. We also propose a few approaches, before acquisition, to either avoid it or correct it at the optical design level.

© 2012 OSA

OCIS Codes
(100.3190) Image processing : Inverse problems
(100.5070) Image processing : Phase retrieval
(100.6890) Image processing : Three-dimensional image processing
(180.2520) Microscopy : Fluorescence microscopy

ToC Category:
Image Processing

Original Manuscript: February 3, 2012
Revised Manuscript: March 30, 2012
Manuscript Accepted: April 3, 2012
Published: April 16, 2012

Virtual Issues
Vol. 7, Iss. 6 Virtual Journal for Biomedical Optics

Praveen Pankajakshan, Zvi Kam, Alain Dieterlen, and Jean-Christophe Olivo-Marin, "Characterizing the 3-D field distortions in low numerical aperture fluorescence zooming microscope," Opt. Express 20, 9876-9889 (2012)

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