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High-resolution 3D optical microscopy inside the beating zebrafish heart using prospective optical gating
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 na...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Optical Society of America
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3521314/ https://www.ncbi.nlm.nih.gov/pubmed/23243558 http://dx.doi.org/10.1364/BOE.3.003043 |
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author | Taylor, Jonathan M. Girkin, John M. Love, Gordon D. |
author_facet | Taylor, Jonathan M. Girkin, John M. Love, Gordon D. |
author_sort | Taylor, Jonathan M. |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-3521314 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Optical Society of America |
record_format | MEDLINE/PubMed |
spelling | pubmed-35213142012-12-14 High-resolution 3D optical microscopy inside the beating zebrafish heart using prospective optical gating Taylor, Jonathan M. Girkin, John M. Love, Gordon D. Biomed Opt Express Microscopy 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. Optical Society of America 2012-10-31 /pmc/articles/PMC3521314/ /pubmed/23243558 http://dx.doi.org/10.1364/BOE.3.003043 Text en © 2012 Optical Society of America http://creativecommons.org/licenses/by-nc-nd/3.0 This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 Unported License, which permits download and redistribution, provided that the original work is properly cited. This license restricts the article from being modified or used commercially. |
spellingShingle | Microscopy Taylor, Jonathan M. Girkin, John M. Love, Gordon D. High-resolution 3D optical microscopy inside the beating zebrafish heart using prospective optical gating |
title | High-resolution 3D optical microscopy inside the beating zebrafish heart using
prospective optical gating |
title_full | High-resolution 3D optical microscopy inside the beating zebrafish heart using
prospective optical gating |
title_fullStr | High-resolution 3D optical microscopy inside the beating zebrafish heart using
prospective optical gating |
title_full_unstemmed | High-resolution 3D optical microscopy inside the beating zebrafish heart using
prospective optical gating |
title_short | High-resolution 3D optical microscopy inside the beating zebrafish heart using
prospective optical gating |
title_sort | high-resolution 3d optical microscopy inside the beating zebrafish heart using
prospective optical gating |
topic | Microscopy |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3521314/ https://www.ncbi.nlm.nih.gov/pubmed/23243558 http://dx.doi.org/10.1364/BOE.3.003043 |
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