Abstract
Contactless sample confinement would enable a whole host of new studies in developmental biology and neuroscience, in particular, when combined with long-term, wide-field optical imaging. To achieve this goal, we demonstrate a contactless acoustic gradient force trap for sample confinement in light sheet microscopy. Our approach allows the integration of real-time environmentally controlled experiments with wide-field low photo-toxic imaging, which we demonstrate on a variety of marine animal embryos and larvae. To illustrate the key advantages of our approach, we provide quantitative data for the dynamic response of the heartbeat of zebrafish larvae to verapamil and norepinephrine, which are known to affect cardiovascular function. Optical flow analysis allows us to explore the cardiac cycle of the zebrafish and determine the changes in contractile volume within the heart. Overcoming the restrictions of sample immobilisation and mounting can open up a broad range of studies, with real-time drug-based assays and biomechanical analyses.
Highlights
Contactless sample confinement would enable a whole host of new studies in developmental biology and neuroscience, in particular, when combined with long-term, wide-field optical imaging
A major advance for realising such studies would be contactless sample suspension combined with rapid and minimally photo-toxic wide-field imaging. This would inherently maintain the integrity of the natural physiological development of the specimen, which may be adversely affected when using agarose or other gels for specimen embedding[1,2,3]
For rapid wide-field imaging, light sheet fluorescence microscopy[4] (LSFM) is becoming established for the advantages it brings to such longitudinal imaging studies
Summary
Contactless sample confinement would enable a whole host of new studies in developmental biology and neuroscience, in particular, when combined with long-term, wide-field optical imaging To achieve this goal, we demonstrate a contactless acoustic gradient force trap for sample confinement in light sheet microscopy. A major advance for realising such studies would be contactless sample suspension combined with rapid and minimally photo-toxic wide-field imaging This would inherently maintain the integrity of the natural physiological development of the specimen, which may be adversely affected when using agarose or other gels for specimen embedding[1,2,3]. A major advance would be a method by which samples could be immobilised in their native environment in the absence of agarose or similar methods of restriction, thereby ensuring more realistic and informative biomedical studies This would enable rapid access of experimental agents in the media to the sample and open new routes for real-time drug assays. Such studies open up the prospect of original avenues in cardiovascular research, long-term imaging and the assessment of dynamic biological processes
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