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<b>Background/Objectives</b>: Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD) is a global health crisis, but current diagnostics are limited. Liver biopsy is invasive, magnetic resonance imaging-proton density fat fraction (MRI-PDFF) is expensive, and quantitative ultrasound methods are low-accuracy, especially in patients with a high body mass index (BMI). This study introduces a novel thermo-acoustic (TA) method that generates ultrasound signals based on tissue electrical conductivity, where lean tissue (high in water and electrolytes) absorbs more radio-frequency (RF) energy than fatty tissue, providing a direct molecular contrast for fat. <b>Methods</b>: A prospective, cross-sectional feasibility study compared a new thermo-acoustic fat fraction (TAFF) score with the reference standard MRI-PDFF in 40 subjects with suspected fatty liver disease. Bland-Altman analysis, Deming regression, and Binary classification performance were tested. To establish system stability, a dedicated Repeatability and Reproducibility (R&R) study (N = 14) evaluated inter-operator and intra-operator consistency using an Intraclass Correlation Coefficient (ICC) derived from a two-way random-effects ANOVA model. <b>Results:</b> TAFF estimates demonstrated a substantial correlation (r = 0.89) with MRI-PDFF and an average absolute error of 3.04% fat fraction. Classification performance was high, with an Area Under the Receiver Operating Characteristic Curve (AUROC) of 0.92 at the 12% fat fraction threshold and 0.99 at the 20% fat fraction threshold. The R&R study confirmed robust stability (intraclass correlation = 0.89) and a negligible mean inter-operator difference of 0.36%. Estimation errors showed no statistically significant correlation with BMI or other body habitus measurements. <b>Conclusions</b>: These findings support thermoacoustics' potential as an accurate, non-invasive, point-of-care solution that can serve as a new imaging biomarker. By providing predictive values closely aligned with MRI-PDFF across the full MASLD spectrum, TAFF can complement currently available ultrasound methods to address the cost and access constraints of MRI for the assessment, diagnosis, and monitoring of MASLD.