Following euthanasia using tricaine (300 mg/L) on ice, whole zebrafish were fixed in 4% paraformaldehyde. For a detailed anatomical study, eye sections were prepared by embedding the samples in paraffin and sectioning through the central region of the eye. These sections were stained with a 0.1% toluidine blue solution containing glacial acetic acid (G3668, Solarbio, China) for visualizing the retinal structure. Microscopic images of these sections were captured at 40 times magnification using a confocal microscope (Olympus FV1000).
For immunofluorescent staining, the sections were treated with an anti-HuC antibody (Cat# 55047-1-AP; Proteintech, China) and an anti-Nat10 antibody (Cat#: 13365–1-AP; Proteintech, China). The stained slides were then analyzed using ImageJ software (developed by Wayne Rasband, National Institutes of Health, Bethesda, MD, USA) to quantify and assess specific histological features.
For immunofluorescent staining, the sections were treated with an anti-HuC antibody (Cat# 55047-1-AP; Proteintech, China) and an anti-Nat10 antibody (Cat#: 13365–1-AP; Proteintech, China). The stained slides were then analyzed using ImageJ software (developed by Wayne Rasband, National Institutes of Health, Bethesda, MD, USA) to quantify and assess specific histological features.