7 December 2013 A high-throughput lab-on-a-chip interface for zebrafish embryo tests in drug discovery and ecotoxicology
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Abstract
Drug discovery screenings performed on zebrafish embryos mirror with a high level of accuracy. The tests usually performed on mammalian animal models, and the fish embryo toxicity assay (FET) is one of the most promising alternative approaches to acute ecotoxicity testing with adult fish. Notwithstanding this, conventional methods utilising 96-well microtiter plates and manual dispensing of fish embryos are very time-consuming. They rely on laborious and iterative manual pipetting that is a main source of analytical errors and low throughput. In this work, we present development of a miniaturised and high-throughput Lab-on-a-Chip (LOC) platform for automation of FET assays. The 3D high-density LOC array was fabricated in poly-methyl methacrylate (PMMA) transparent thermoplastic using infrared laser micromachining while the off-chip interfaces were fabricated using additive manufacturing processes (FDM and SLA). The system’s design facilitates rapid loading and immobilization of a large number of embryos in predefined clusters of traps during continuous microperfusion of drugs/toxins. It has been conceptually designed to seamlessly interface with both upright and inverted fluorescent imaging systems and also to directly interface with conventional microtiter plate readers that accept 96-well plates. We also present proof-of-concept interfacing with a high-speed imaging cytometer Plate RUNNER HD® capable of multispectral image acquisition with resolution of up to 8192 x 8192 pixels and depth of field of about 40 μm. Furthermore, we developed a miniaturized and self-contained analytical device interfaced with a miniaturized USB microscope. This system modification is capable of performing rapid imaging of multiple embryos at a low resolution for drug toxicity analysis.
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Feng Zhu, Feng Zhu, Jin Akagi, Jin Akagi, Chris J. Hall, Chris J. Hall, Kathryn E. Crosier, Kathryn E. Crosier, Philip S. Crosier, Philip S. Crosier, Pierre Delaage, Pierre Delaage, Donald Wlodkowic, Donald Wlodkowic, } "A high-throughput lab-on-a-chip interface for zebrafish embryo tests in drug discovery and ecotoxicology", Proc. SPIE 8923, Micro/Nano Materials, Devices, and Systems, 892345 (7 December 2013); doi: 10.1117/12.2033404; https://doi.org/10.1117/12.2033404
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