Embryonic and larvae zebrafish on microfluidic chip.

(A) Schematic of chip for zebrafish assay. The chip includes two independent zones, each with a media inlet, a drug inlet, a gradient generator and seven series of fish tanks (one concentration with three tanks) named C1–C7. Left zone for embryonic toxicity and teratogenicity experiment, the right for larvae fish based drug toxicity evaluation. (B) Photos of the microfluidic chip and micrographs of the embryo and larvae in the chip.

Development of Zebrafish embryos in microfluidic chip under flowing stream.

(A) Hatching percentage of embryos cultured in the chip by continuous exposure at varied flow rates and at different growth periods. The asterisks indicate significant differences from control group (chamber 1) * at p<0.05. (B) The body length of the larvae fish at 96 hpf (n = 20) cultured in the chip by continuous exposure at varied flow rates (pink control is body length of the larvae fish cultured in 24 microwells).

Apl induced abnormal morphology, developmental retardation, and mortality of zebrafish embryos.

(A) Hatched rate and (B) mortality of zebrafish embryos exposed to gradient Apl every 12 hpf in the microfluidic chip for 96 h. (C) Typical morphological abnormalities of embryos exposed to Apl. Red arrows indicate tail malformation, delayed yolk absorption, pericardial edema, and bent trunk, respectively (from upper left-right to bottom left-right). (D) Mean body length of hatched embryos treated with Apl in the chip compared with the controls (C1) at 72 hpf (n = 6), there are no date at C6 and C7 for the fish are not hatched. The asterisks indicate significant differences from control group (chamber 1) * at p<0.05.