TY  - JOUR
TI  - Computational Quantification of Mouse Retinal Vasculature Using ImageJ
AU  - Nader, Michel 
AU  - Feridooni, Hirad A.
AU  - Tavasoli, Mahtab 
AU  - Van Der Ende, Sarah 
AU  - McMaster, Christopher R.
AU  - Robitaille, Johane M.
VL  - 16
IS  - 11
PY  - 2026
DA  - 2026/06/05
SP  - e5705
C1  - Bio-protocol 2026;16:e5705
DO  - 10.21769/BioProtoc.5705
UR  - https://doi.org/10.21769/BioProtoc.5705
AB  - Postnatal mouse retinal vascular development is a widely used model for studying retinal vascular diseases and evaluating candidate therapies. This is particularly relevant for inherited disorders such as familial exudative vitreoretinopathy (FEVR), in which impaired vascular growth and organization are central to disease pathogenesis. Numerous approaches have been used to assess retinal vasculature in mouse flat mounts, ranging from qualitative descriptions to limited quantitative measurements of vascular growth. However, phenotypic variability across genetic models, including different models of FEVR, complicates comparisons and underscores the need for standardized, comprehensive multi-parameter analyses that are suitable for rapid and cost-effective screening studies. We describe a standardized morphometric protocol using ImageJ software to quantitatively analyze mouse retinal vasculature in a reproducible manner. The protocol begins with measurement of areas of vascular disorganization (meshes) as well as total vascular and retinal area. Two defined regions in the peripheral and midperipheral retina are then selected to quantify cell clusters, followed by image processing, binarization, and skeletonization. From these processed images, vascular density, branch number, branch length and thickness, junction number, triple points, and box-counting fractal dimension and lacunarity are quantified. Overall, this protocol provides a rapid, cost-effective, and standardized framework for quantifying retinal vascular phenotypes across diverse mouse models. By capturing multiple structural features and accommodating phenotypic variability, it is well-suited for comparative studies and therapeutic screening in retinal vascular disease.
KW  - Retina
KW  - Vessels
KW  - Mice
KW  - Screening tool
KW  - Disease models
KW  - Candidate treatments
KW  - Image analysis
KW  - ImageJ
KW  - FIJI
KW  - Fractal analysis
KW  - Image processing
KW  - Morphometry
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False