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Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 3, Iss. 12 — Dec. 1, 2012
  • pp: 3043–3053

High-resolution 3D optical microscopy inside the beating zebrafish heart using prospective optical gating

Jonathan M. Taylor, John M. Girkin, and Gordon D. Love  »View Author Affiliations

Biomedical Optics Express, Vol. 3, Issue 12, pp. 3043-3053 (2012)

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3D fluorescence imaging is a fundamental tool in the study of functional and developmental biology, but effective imaging is particularly difficult in moving structures such as the beating heart. We have developed a non-invasive real-time optical gating system that is able to exploit the periodic nature of the motion to acquire high resolution 3D images of the normally-beating zebrafish heart without any unnecessary exposure of the sample to harmful excitation light. In order for the image stack to be artefact-free, it is essential to use a synchronization source that is invariant as the sample is scanned in 3D. We therefore describe a scheme whereby fluorescence image slices are scanned through the sample while a brightfield camera sharing the same objective lens is maintained at a fixed focus, with correction of sample drift also included. This enables us to maintain, throughout an extended 3D volume, the same standard of synchronization we have previously demonstrated in and near a single 2D plane. Thus we are able image the complete beating zebrafish heart exactly as if the heart had been artificially stopped, but sidestepping this undesirable interference with the heart and instead allowing the heart to beat as normal.

© 2012 OSA

OCIS Codes
(110.2960) Imaging systems : Image analysis
(110.6880) Imaging systems : Three-dimensional image acquisition
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(170.3880) Medical optics and biotechnology : Medical and biological imaging
(180.6900) Microscopy : Three-dimensional microscopy

ToC Category:

Original Manuscript: August 15, 2012
Revised Manuscript: September 25, 2012
Manuscript Accepted: October 20, 2012
Published: October 31, 2012

Jonathan M. Taylor, John M. Girkin, and Gordon D. Love, "High-resolution 3D optical microscopy inside the beating zebrafish heart using prospective optical gating," Biomed. Opt. Express 3, 3043-3053 (2012)

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