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<p>Quantification of retinal and avascular areas in the rat oxygen induced retinopathy (OIR) model is traditionally performed using time-intensive manual methods that may be subject to inter-rater variability. We developed and validated two open-source, semi-automated Fiji (Fiji is Just ImageJ) macros to measure total retinal area (RA) and avascular area (AA) in rat 50/10 OIR retinal flat mounts. Concordance correlation coefficients demonstrated high agreement between macro-derived and manual measurements for RA (0.988), AA (0.985), and AA/RA (0.983), with minimal bias observed. While inter-user variability was higher for macro-based AA measurements compared to manual methods, averaging results from two users improved precision. These semi-automated tools reduce analysis time and training burden while maintaining strong agreement with gold standard measurements. Although limited by user input requirements and single-laboratory validation, these macros provide accessible, standardized methods for retinal quantification and have the potential to improve reproducibility and efficiency in OIR research.</p>

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Semi-automated quantification of total and avascular retinal area in the rat model of oxygen-induced retinopathy

  • 期刊论文

  • 发表日期：

    2026年8月1日

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  • Husaam Qureishy,
  • Amelia Morikuni,
  • Michael D. Evans,
  • Rehan Krishnan,
  • Ellen C. Ingolfsland

摘要

Quantification of retinal and avascular areas in the rat oxygen induced retinopathy (OIR) model is traditionally performed using time-intensive manual methods that may be subject to inter-rater variability. We developed and validated two open-source, semi-automated Fiji (Fiji is Just ImageJ) macros to measure total retinal area (RA) and avascular area (AA) in rat 50/10 OIR retinal flat mounts. Concordance correlation coefficients demonstrated high agreement between macro-derived and manual measurements for RA (0.988), AA (0.985), and AA/RA (0.983), with minimal bias observed. While inter-user variability was higher for macro-based AA measurements compared to manual methods, averaging results from two users improved precision. These semi-automated tools reduce analysis time and training burden while maintaining strong agreement with gold standard measurements. Although limited by user input requirements and single-laboratory validation, these macros provide accessible, standardized methods for retinal quantification and have the potential to improve reproducibility and efficiency in OIR research.