A Bubble-Free Microfluidic Device for Easy-to-Operate Immobilization, Culturing and Monitoring of Zebrafish Embryos

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.

Lateral Degassing Method for Disposable Film-Chip Microfluidic Devices

“…Meanwhile, PDMS is a commonly used material for microfluidic device fabrication [ 27 ] and is also known for its high gas permeability [ 28 , 29 ]. Therefore, a variety of active degassing methods using PDMS membranes have been developed for a wide range of microfluidic applications, such as PCR [ 30 ], cell culture [ 31 ], drug screening [ 32 ], and fuel cells [ 33 ]. Most PDMS-based active degassing methods use vacuum microchambers for bubble extraction [ 34 , 35 ], which is usually implemented in a multi-layered structure, making device fabrication difficult and impractical.…”

Cost-effective fabrication of photopolymer molds with multi-level microstructures for PDMS microfluidic device manufacture

“…27 Thus, the fabrication of molds with multilevel microstructures for the PDMS microdevice fabrication presents a useful potential for biological applications. 18,20,28,29 Recently, we reported the manufacture of PDMS microuidic devices by employing a exographic photopolymer as mold. 30,31 Advantages related to resolution, mold size, aspect ratio, roughness, availability, scalability and costs were demonstrated.…”

3D-Printed Bubble-Free Perfusion Cartridge System for Live-Cell Imaging

“…It has millimeter-scale bent and flat tapered flow channels that prevent air bubbles from entering the observation area without the need for additional bubble-removing devices. Various bubble trap systems based on microfluidic technologies have been proposed previously [ 11 , 12 , 13 , 14 , 15 , 16 ]. For instance, researchers have developed a two-layer bubble barrier structure, in which the top layer blocks bubbles and the bottom layer works as a fluidic channel [ 11 ], and a debubbler using a polydimethylsiloxane (PDMS) pneumatic layer [ 12 ].…”