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The goal of this study was to evaluate the magnitude and origin of scattered radiation in a small-animal PET scanner and to assess the impact of these findings on noise equivalent count rate (NECR) measurements, a metric often used to optimize scanner acquisition parameters and to compare one scanner with another. The scatter fraction (SF) was measured for line sources in air and line sources placed within a mouse-sized phantom (25 mm φ×70 mm) and a rat-sized phantom (60 mm φ×150 mm) on the microPET II small-animal PET scanner. Measurements were performed for lower energy thresholds ranging from 150-450 keV and a fixed upper energy threshold of 750 keV. Four different methods were compared for estimating the SF. Significant scatter fractions were measured with just the line source in the field of view, with the spatial distribution of these events consistent with scatter from the gantry and room environment. For mouse imaging, this component dominates over object scatter, and the measured SF is strongly method dependent. The environmental SF rapidly increases as the lower energy threshold decreases and can be more than 30% for an open energy window of 150-750 keV. The object SF originating from the mouse phantom is about 3-4% and does not change significantly as the lower energy threshold increases. The object SF for the rat phantom ranges from 10 to 35% for different energy windows and increases as the lower energy threshold decreases. Because the measured SF is highly dependent on the method, and there is as yet no agreed upon standard for animal PET, care must be exercised when comparing NECR for small objects between different scanners. Differences may be methodological rather than reflecting any relevant difference in the performance of the scanner. Furthermore, these results have implications for scatter correction methods when the majority of the detected scatter does not arise from the object itself.