Exploring discrepancies in muscle analysis with ImageJ: understanding the impact of tool selection on echo intensity and muscle area measurements

The aim was to compare the use of different tools within the ImageJ program (polygon vs. segmented line) and their impact on the calculation of muscle area and echo intensity (EI) values in ultrasound imaging of the vastus lateralis muscle.

Thirteen volunteers participated in this study. Ultrasound images of the vastus lateralis muscle were acquired using 2D B-mode ultrasonography and analyzed using both the polygon and segmented line tools by the same evaluator. The intraclass correlation coefficient (ICC) and coefficient of variation (CV) assessed the tools' reliability. Bland–Altman plots were employed to verify the agreement between measurements, and linear regression analysis determined proportional bias. A paired t-test was conducted to analyze differences between the tools.

The reliability between tools for muscle area calculation was weak (r = 0.000; CV = 138.03 ± 0.34%), while it was excellent for EI (r = 0.871; CV = 15.19 ± 2.96%). The Bland–Altman plots indicated a large bias for muscle area (d = 195.2%) with a proportional bias (p < 0.001). For EI, the bias was (d = 15.2) with proportional bias (p = 0.028). The paired t-test revealed significant differences between the tools for area (p < 0.001) but not for EI (p = 0.060).

The study found significant differences in measurements obtained with the polygon and segmented line tools in ImageJ, with the polygon tool showing higher values for muscle area and lower values for EI.