9 March 2013 Dynamic analysis of angiogenesis in transgenic zebrafish embryos using a 3D multilayer chip-based technology
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Abstract
Transgenic zebrafish (Danio rerio) models of human diseases have recently emerged as innovative experimental systems in drug discovery and molecular pathology. None of the currently available technologies, however, allow for automated immobilization and treatment of large numbers of spatially encoded transgenic embryos during real-time developmental analysis. This work describes the proof-of-concept design and validation of an integrated 3D microfluidic chip-based system fabricated directly in the poly(methyl methacrylate) transparent thermoplastic using infrared laser micromachining. At its core, the device utilizes an array of 3D micro-mechanical traps to actively capture and immobilize single embryos using a low-pressure suction. It also features built-in piezoelectric microdiaphragm pumps, embryo trapping suction manifold, drug delivery manifold and optically transparent indium tin oxide (ITO) heating element to provide optimal temperature during embryo development. Furthermore, we present design of the proof-of-concept off-chip electronic interface equipped with robotic servo actuator driven stage, innovative servomotor-actuated pinch valves and miniaturized fluorescent USB microscope. Our results show that the innovative device has 100% embryo trapping efficiency while supporting normal embryo development for up to 72 hours in a confined microfluidic environment. We also present data that this microfluidic system can be readily applied to kinetic analysis of a panel of investigational anti-angiogenic agents in transgenic zebrafish Tg(fli1a:EGFP) line. The optical transparency and embryo immobilization allow for convenient visualization of developing vasculature patterns in response to drug treatment without the need for specimen re-positioning. The integrated electronic interfaces bring the Lab-on-a-Chip systems a step closer to realization of complete analytical automation.
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Jin Akagi, Jin Akagi, Feng Zhu, Feng Zhu, Chris J. Hall, Chris J. Hall, Khashayar Khoshmanesh, Khashayar Khoshmanesh, Kourosh Kalantar-Zadeh, Kourosh Kalantar-Zadeh, Arnan Mitchell, Arnan Mitchell, Kathryn E. Crosier, Kathryn E. Crosier, Philip S. Crosier, Philip S. Crosier, Donald Wlodkowic, Donald Wlodkowic, } "Dynamic analysis of angiogenesis in transgenic zebrafish embryos using a 3D multilayer chip-based technology", Proc. SPIE 8615, Microfluidics, BioMEMS, and Medical Microsystems XI, 86151B (9 March 2013); doi: 10.1117/12.2001467; https://doi.org/10.1117/12.2001467
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