Home › Cell Biology › Studying Diabetes Through the Eyes of a Fish: Microdissection, Visualization, and Analysis of the Adult tg(fli:EGFP) Zebrafish Retinal Vasculature
Cell Biology JoVE (Open Access) Citable · DOI

Studying Diabetes Through the Eyes of a Fish: Microdissection, Visualization, and Analysis of the Adult tg(fli:EGFP) Zebrafish Retinal Vasculature

DOI: 10.3791/56674-v
What you'll learn
  • ✓ Perform microdissection and tissue preparation of adult zebrafish retinas
  • ✓ Mount and visualize fluorescent retinal vasculature using microscopy
  • ✓ Analyze vascular structure and neoangiogenesis in disease models
  • ✓ Quantify morphological changes in transgenic zebrafish vasculature
Protocol

Here, we discuss a method protocol which will allow an easy analysis of the adult tg(fli:EGFP) zebrafish retinal vasculature as a fast read-out in settings of long-term vascular pathologies linked to neoangiogenesis and structural changes.

Difficulty
intermediate
Total time
~2–3 hours per zebrafish
Model organism
Zebrafish tg(fli:EGFP)

Steps

1
Fix zebrafish tissue with paraformaldehyde

Euthanize and perfuse adult tg(fli:EGFP) zebrafish, then fix eyes in paraformaldehyde solution to preserve tissue structure and fluorescent protein signal.

▶ 00:54
2
Microdissect retinal vasculature from eye

Carefully remove and isolate the retinal tissue from fixed eyes, separating vascular structures for downstream analysis and mounting.

▶ 01:41
3
Mount retina and prepare for visualization

Arrange dissected retinal tissue on microscope slides with mounting medium to optimize optical clarity and fluorescent signal visualization.

▶ 06:39
4
Analyze retinal vascular morphology and structure

Use fluorescence microscopy to image the tg(fli:EGFP) retinal vasculature and quantify vascular parameters such as branching, density, and neoangiogenic changes.

▶ 07:51
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