Hedde Per Niklas, Le Brian T, Gomez Erika L, Duong Leora, Steele Robert E, Ahrar Siavash
Beckman Laser Institute and Medical Clinic, University of California Irvine, Irvine, CA 92612, USA.
Department of Biomedical Engineering, CSU Long Beach, Long Beach, CA 90840, USA.
Biology (Basel). 2023 Jan 11;12(1):116. doi: 10.3390/biology12010116.
Selective plane illumination microscopy (SPIM), or light sheet microscopy, is a powerful imaging approach. However, access to and interfacing microscopes with microfluidics have remained challenging. Complex interfacing with microfluidics has limited the SPIM's utility for studying the hydrodynamics of freely moving multicellular organisms. We developed SPIM-Flow, an inexpensive light sheet platform that enables easy integration with microfluidics. We used SPIM-Flow to investigate the hydrodynamics of a freely moving polyp via particle tracking in millimeter-sized chambers. Initial experiments across multiple animals, feeding on a chip ( nauplii used as food), and baseline behaviors (tentacle swaying, elongation, and bending) indicated the organisms' health inside the system. Fluidics were used to investigate 's response to flow. The results suggested that the animals responded to an established flow by bending and swaying their tentacles in the flow direction. Finally, using SPIM-Flow in a proof-of-concept experiment, the shear stress required to detach an animal from a surface was demonstrated. Our results demonstrated SPIM-Flow's utility for investigating the hydrodynamics of freely moving animals.
选择性平面照明显微镜(SPIM),即光片显微镜,是一种强大的成像方法。然而,将显微镜与微流体装置连接并使用一直具有挑战性。与微流体装置的复杂连接限制了SPIM在研究自由移动的多细胞生物流体动力学方面的应用。我们开发了SPIM-Flow,这是一种低成本的光片平台,能够轻松与微流体装置集成。我们使用SPIM-Flow通过在毫米级腔室中进行粒子追踪来研究自由移动的水螅的流体动力学。对多只动物进行的初步实验,包括在芯片上进食(以无节幼体作为食物)以及基线行为(触手摆动、伸长和弯曲),表明了系统内生物的健康状况。利用流体学研究了动物对水流的反应。结果表明,动物会通过在水流方向上弯曲和摆动触手来对既定水流做出反应。最后,在一个概念验证实验中使用SPIM-Flow,证明了将动物从表面分离所需的剪切应力。我们的结果证明了SPIM-Flow在研究自由移动动物流体动力学方面的实用性。