X cite 120 ledmini

Spinal neurons were counted in Tg(isl1:GFP) and Tg(chx10:GFP) 3-dpf larvae. For wholemount imaging of Tg(chx10:GFP) larvae, animals were anesthetized in 0.02% tricaine and pinned laterally through the notochord in a Sylgard coated dish. Imaging was completed using a Zeiss Examiner.D1 microscope equipped with an Axiocam 506 mono Zeiss camera and an X-Cite 120 LEDmini (Excelitas Technologies) light excitation source for fluorescence; 100-μm deep Z-stacks were performed using Zen pro software, and we performed cell counts manually.
Immunostained spinal cord sections of Tg(isl1:GFP) larvae and brain sections of Tg(dat:CFP-NTR) larvae were imaged using an Olympus FV1000 spectral LSM confocal microscope with Z-stack intervals of 0.5 μm. Imaging was performed at 20× and 40× magnifications, with a resolution of 1024 × 10²⁴ pixels. We performed three to four independent counts for each Z-stack using Fluoview or ImageJ, and means were computed.

The ROS content was examined using a fluorescent probe, 2′-7′dichlorofluorescin diacetate (DCFH-DA; Sigma Aldrich, St. Louis, MO, USA) in live zebrafish larvae at 48, 72, and 96 hpf. Zebrafish larvae were briefly rinsed twice with PBS and incubated with 3 μM DCFH-DA for 2 h in the dark at RT and then washed three times with PBS for 5 min each. The lateral view whole zebrafish body images of each embryo were captured using a microscopic camera (Amscope HDMI Color CMOS C-mount Camera, Amscope, Irvine, CA, USA) attached to an Olympus SZ51 stereomicroscope (Olympus, Barrington, NJ, USA), illuminated by NIGHTSEA (Lexington, MA, USA) fluorescence illumination system with a royal blue excitation cube. For higher magnification (3×) focusing on the head only, each live zebrafish larvae were mounted with low-melt agarose (1.5%) onto the depressed microscopic slide. The ventral/dorsal view images of each larva were captured using an Amscope Digital camera (5MP USB3.0 High-Speed Color CMOS C-mount Microscope Camera, Amscope, Irvine, CA, USA) attached to an Olympus SZX7 stereomicroscope (Olympus, Barrington, NJ, USA), illuminated by X-Cite120 LED mini (Excelitas Technologies, Waltham, MA, USA) with the fluorescence light power between 8 and 12%. The captured ROS images were analyzed by imaging software FIJI. Different density was measured and presented as a fold change compared to control.