Abstract
The development of miniaturized devices for studying zebrafish embryos has been limited due to complicated fabrication and operation processes. Here, we reported on a microfluidic device that enabled the capture and culture of zebrafish embryos and real-time monitoring of dynamic embryonic development. The device was simply fabricated by bonding two layers of polydimethylsiloxane (PDMS) structures replicated from three-dimensional (3D) printed reusable molds onto a flat glass substrate. Embryos were easily loaded into the device with a pipette, docked in traps by gravity, and then retained in traps with hydrodynamic forces for long-term culturing. A degassing chamber bonded on top was used to remove air bubbles from the embryo-culturing channel and traps so that any embryo movement caused by air bubbles was eliminated during live imaging. Computational fluid dynamics simulations suggested this embryo-trapping and -retention regime to exert low shear stress on the immobilized embryos. Monitoring of the zebrafish embryogenesis over 20 h during the early stages successfully verified the performance of the microfluidic device for culturing the immobilized zebrafish embryos. Therefore, this rapid-prototyping, low-cost and easy-to-operate microfluidic device offers a promising platform for the long-term culturing of immobilized zebrafish embryos under continuous medium perfusion and the high-quality screening of the developmental dynamics.
Highlights
The zebrafish, Danio rerio, has become a prominent vertebrate model for disease modeling and drug discovery [1]
With the development of advanced manufacturing technologies such as microfabrication, laser micromachining and three-dimensional (3D) printing, dedicated microfluidic devices for the immobilization, flow-perfusion culture, dosing and time-lapse imaging of zebrafish embryos have emerged in the past decade [9,10,11]
During the long-term culturing and monitoring of zebrafish embryos, any air bubbles brought to the embryo-culturing channel and traps may squeeze the immobilized embryo out of its original position or obstruct the view of imaging, and affect the stability of embryo immobilization and the quality of time-lapse imaging
Summary
The zebrafish, Danio rerio, has become a prominent vertebrate model for disease modeling and drug discovery [1]. With the development of advanced manufacturing technologies such as microfabrication, laser micromachining and three-dimensional (3D) printing, dedicated microfluidic devices for the immobilization, flow-perfusion culture, dosing and time-lapse imaging of zebrafish embryos have emerged in the past decade [9,10,11]. Zhu et al developed a 3D high-throughput microfluidic platform, which enables the stable immobilization of single embryos by combining continuous medium perfusion at a flow rate of 400 μL/min and aspiration via horizontal tunnels embedded between the traps and suction channel [17].
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