High-resolution 3D optical microscopy inside the beating zebrafish heart using prospective optical gating
| Autor: | Jonathan M. Taylor, John M. Girkin, Gordon D. Love |
|---|---|
| Rok vydání: | 2012 |
| Předmět: |
Microscopy
Fluorescence-lifetime imaging microscopy biology business.industry Computer science ocis:(170.2520) Fluorescence microscopy Beat (acoustics) High resolution Gating biology.organism_classification Atomic and Molecular Physics and Optics law.invention Optics Optical microscope law High-speed photography ocis:(180.6900) Three-dimensional microscopy ocis:(110.6880) Three-dimensional image acquisition Tomography ocis:(110.2960) Image analysis ocis:(170.3880) Medical and biological imaging business Zebrafish Biotechnology |
| Zdroj: | Biomedical Optics Express |
| ISSN: | 2156-7085 |
| DOI: | 10.1364/boe.3.003043 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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