SUPA, School of Physics and Astronomy, University of St Andrews, St Andrews, KY16 9SS, UK.
Electron Bio-Imaging Centre, Diamond Light Source, Harwell Science and Innovation Campus, Didcot, OX11 0DE, UK.
Nat Commun. 2019 Feb 8;10(1):669. doi: 10.1038/s41467-019-08514-5.
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.
无接触式样品限制将使在发展生物学和神经科学领域进行大量新的研究成为可能，特别是与长期、宽场光学成像结合使用时。为了实现这一目标，我们展示了一种用于光片显微镜中样品限制的无接触式声梯度力陷阱。我们的方法允许将实时环境控制实验与宽场低光毒性成像相结合，我们在各种海洋动物胚胎和幼虫上进行了演示。为了说明我们方法的主要优势，我们提供了定量数据，用于研究已知影响心血管功能的药物维拉帕米和去甲肾上腺素对斑马鱼幼虫心跳的动态反应。光流分析使我们能够探索斑马鱼的心脏周期，并确定心脏内收缩体积的变化。克服样品固定和安装的限制可以开辟广泛的研究领域，包括实时基于药物的测定和生物力学分析。
