BACKGROUND: Alzheimer's disease (AD) is a progressive neurodegenerative disorder that is the leading cause of dementia, characterized by memory loss, cognitive decline, and significant social and economic burdens. Despite extensive research into amyloid positron emission tomography (PET) radiopharmaceuticals, the effectiveness of various F-labeled tracers for imaging amyloid plaques in AD mouse models remains uncertain. This study aimed to evaluate the performance of three radiopharmaceuticals-F-florbetaben (FBB), F-flutemetamol (FMM), and F-florapronol (FPN)-in differentiating amyloid deposition in AD and control mice. RESULTS: F-FMM and F-FBB demonstrated significantly higher standardized uptake value ratios (SUVRs) in AD mice than in controls. For F-FBB, the mean SUVR in AD mice was 1.06, significantly higher than the 0.81 in controls (p < 0.001). Similarly, F-FMM showed a mean SUVR of 0.97 in AD mice compared to 0.94 in controls (p = 0.024). In contrast, F-FPN did not show significant SUVR differences between AD and control groups (p = 0.071). Comparative analysis revealed that F-FBB exhibited a significantly greater SUVR difference between AD and control groups compared to F-FMM (p < 0.001). CONCLUSIONS: F-FBB emerged as the most effective radiopharmaceutical for imaging amyloid deposition in AD mouse models, providing superior differentiation between AD and control groups. These findings support the optimization of amyloid PET tracers for preclinical studies, facilitating advancements in Alzheimer's research. CLINICAL TRIAL NUMBER: Not applicable.