Design and use of PET laminate microfluidics device (RADISH). (A) Schematic illustrating individual PET layers' arrangement and thickness, assembly, and device mounting and use. (B) Final product, mounted in a ¾" cutout in a 35 mm dish. (C) 3 dpf embryos, (D) 5 dpf embryos, (E) 14 dpf embryos housed inside the device. Scale bars = 1,000 µm (C,D,E). Abbreviations: PET = polyethylene terephthalate; RADISH = Rotational Assistant for Danio Imaging of Subsequent Healing; dpf = days post fertilization

Step-by-step photographic illustration of PET device assembly. (A) Individual PET layers before lamination. (B) PET layer secured to 25 mm coverslip using office tape. (C) Device-in-progress secured to a plastic backing for lamination. (D) Completed PET laminate device. (E) Modified 35 mm dish before device mounting. (F) Completed device. Abbreviation: PET = polyethylene terephthalate.

Improved quality of laser scanning confocal images of 3 dpf embryos in the RADISH design. (A) Representative image of 3 dpf Tg(krt4:Gal4, UAS:GCaMP6f) embryo stained with MemGlow 560 without using the RADISH. (B) Representative image of Tg(krt4:Gal4, UAS:GCaMP6f) embryo stained with MemGlow 560 mounted in the RADISH. Scale bar = 100 µm. Abbreviations: RADISH = Rotational Assistant for Danio Imaging of Subsequent Healing; dpf = days post fertilization

Compatibility of the RADISH design with multiple wounding techniques. (A) 3 dpf Tg(krt4:Gal4, UAS:GCaMP6f) x (cdh1-dTomato)xt18 embryo wounded via manual tailfin transection. (B) Embryo wounded via laser ablation. (C,D) The same embryos 4 min later, showing the progression of calcium transients after wounding. Scale bars = 100 µm. Abbreviations: RADISH = Rotational Assistant for Danio Imaging of Subsequent Healing; dpf = days post fertilization

Long-term entrapment of embryos for imaging of phagocyte migration. (A) Tol2(mpx:Dendra2) embryos immediately post wounding, and the same embryos (B) 25 min, (C) 12 h, and (D) 24 h later. Scale bars = 100 µm