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

Taylor, J.M. , Girkin, J.M. and Love, G.D. (2012) High-resolution 3D optical microscopy inside the beating zebrafish heart using prospective optical gating. Biomedical Optics Express, 3(12), pp. 3043-3053. (doi: 10.1364/BOE.3.003043)

Abstract

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.

• Item Type: Articles
• Additional Information: This paper was published in Biomedical Optics Express and is made available as an electronic reprint with the permission of OSA. The paper can be found at the following URL on the OSA website http://dx.doi.org/10.1364/BOE.3.003043. Systematic or multiple reproduction or distribution to multiple locations via electronic or other means is prohibited and is subject to penalties under law.
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Taylor, Dr Jonathan
• Authors: Taylor, J.M., Girkin, J.M., and Love, G.D.
• College/School: College of Science and Engineering > School of Physics and Astronomy
• Journal Name: Biomedical Optics Express
• Publisher: Optical Society of America
• ISSN: 2156-7085
• Published Online: 31 October 2012
• Copyright Holders: Copyright © 2012 Optical Society of America
• First Published: First published in Biomedical Optics Express 3(12):3043-3053
• Publisher Policy: Reproduced in accordance with the copyright policy of the publisher