Abstract
Purpose :
Accurate and precise as well as efficient measurements of choroidal neovascularization (CNV) lesions are important in the laser-induced CNV mouse model, one of the major animal models used in developing therapeutic agents for various neovascular retinal diseases. We present the utility of FIJI-assisted automatic quantification of the total volume of CNV.
Methods :
Laser-induced CNV was induced in C57BL/6J mice according to the established protocol. Following CNV induction, mice were treated with intravitreal injection of either PBS (n=6-7) or aflibercept (Eylea®) (n=11). One week after treatment, RPE/choroid flat mounts were stained with rhodamine-conjugated Griffonia simplicifolia lectin B4 (GSA). Z-stacks of the entire CNV lesion obtained using laser confocal microscopy were converted to binary stacks using FIJI. Thresholded voxels were quantified using the Voxel-Counter Plugin. Data from volumetric analysis and multiple area analyses from blindly selected, mean, and a maximum of measured areas using FIJI were compared.
Results :
FIJI-assisted automatic quantitative volumetric analysis of CNV was useful in detecting experimental outliers in laser-induced CNV genesis and provided accurate and precise measurements of total areas of CNV with a lower coefficient of variance (63%) than in multiple area analyses, including blindly selected, mean, and a maximum of measured areas (76%, 69%, and 67% respectively). A lower coefficient of variance resulted in increased statistical significance (p-value) when comparing CNV lesions in PBS and Eylea treated groups (Figure).
Conclusions :
FIJI-assisted automatic quantitative volumetric analysis can be useful for accurate, precise and efficient measurements of total areas of CNV. Further, this approach can be advantageous in determining the efficacy of therapeutic agents in the laser-induced CNV mouse model.
This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.