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Background.  Quantitative diagnostic ultrasound imaging has been proposed as a method of estimating muscle quality using measures of echogenicity. The Rectangular Marquee Tool (RMT) and the Free Hand Tool (FHT) are two types of editing features used in Photoshop and ImageJ for determining a region of interest (ROI) within an ultrasound image. The primary objective of this study is to determine the intrarater and interrater reliability of Photoshop and ImageJ for the estimate of muscle tissue echogenicity in older adults via grayscale histogram analysis. The secondary objective is to compare the mean grayscale values obtained using both the RMT and FHT methods across both image analysis platforms.   Methods.  This cross-sectional observational study features 18 community-dwelling men (age = 61.5 ± 2.32 years). Longitudinal views of the rectus femoris were captured using B-mode ultrasound. The ROI for each scan was selected by 2 examiners using the RMT and FHT methods from each software program. Their reliability is assessed using intraclass correlation coefficients (ICCs) and the standard error of the measurement (SEM). Measurement agreement for these values is depicted using Bland-Altman plots. A paired  t -test is used to determine mean differences in echogenicity expressed as grayscale values using the RMT and FHT methods to select the post-image acquisition ROI. The degree of association among ROI selection methods and image analysis platforms is analyzed using the coefficient of determination ( R  2 ).   Results.  The raters demonstrated excellent intrarater and interrater reliability using the RMT and FHT methods across both platforms (lower bound 95% CI ICC = .97–.99,  p  < .001). Mean differences between the echogenicity estimates obtained with the RMT and FHT methods was .87 grayscale levels (95% CI [.54–1.21],  p  < .0001) using data obtained with both programs. The SEM for Photoshop was .97 and 1.05 grayscale levels when using the RMT and FHT ROI selection methods, respectively. Comparatively, the SEM values were .72 and .81 grayscale levels, respectively, when using the RMT and FHT ROI selection methods in ImageJ. Uniform coefficients of determination ( R  2  = .96–.99,  p  < .001) indicate strong positive associations among the grayscale histogram analysis measurement conditions independent of the ROI selection methods and imaging platform.   Conclusion.  Our method for evaluating muscle echogenicity demonstrated a high degree of intrarater and interrater reliability using both the RMT and FHT methods across 2 common image analysis platforms. The minimal measurement error exhibited by the examiners demonstrates that the ROI selection methods used with Photoshop and ImageJ are suitable for the post-acquisition image analysis of tissue echogenicity in older adults.

peerj

Ultrasound estimates of muscle quality in older adults: reliability and comparison of Photoshop and ImageJ for the grayscale analysis of muscle echogenicity