Abstract:

Ｏｂｊｅｃｔｉｖｅ：Zebrafish has emerged as a vertebrate model system for gene function studying and drug screening. Zebrafish phenotype is usually presented by acquiring and analyzing images of the dorsal and lateral view of zebrafish embryos. In this study, we utilized desktop 3D printers to fabricate 96well plate and dish compatible orientation tools for observing the phenotype of zebrafish podocytes after injury.
Ｍｅｔｈｏｄｏｌｏｇｙ：Transgenic zebrafish Tg (pod; Gal4; UAS: NTRmCherry) was generated as inducible podocyte injury model. Induction of podocyte injury by metronidazole (MTZ), which was converted to a cytotoxin by the bacterial nitroreductase (NTR) transgenically expressed in zebrafish podocytes, led to glomerular filtration barrier (GFB) damage. 3D objects were designed using OpenSCAD software and printed on Object 500 3D printer. Results: The printed tools were used to generate deep agarose cavities allowing a fixed anteroposterior orientation and reduce movement of embryo. The lateral and dorsal views of the zebrafish embryos arranged in the microcavity were acquired by automatic microscopy for observing edema and fluorescence.
Ｒｅｓｕｌｔｓ：The printed tools were used to generate deep agarose cavities allowing a fixed anteroposteriororientation and reduce movement of embryo.The lateral and dorsal views of the zebrafish embryos arranged in the microcavity were acquired by automatic microscopy for observing edema and fluorescence.
Ｃｏｎｃｌｕｓｉｏｎ：We demonstrate the utilization of desktop 3D printers to fabricate orientation tools for zebrafish embryos enabling the acquisition of consistent lateral or dorsal views in screening assays。

Key words: zebrafish screening, high content screening, 3D printingpodocyte, proteinuria