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Objective Dynamic positron emission tomography (PET), which reveals information about both the spatial distribution and temporal kinetics of a radiotracer, enables quantitative interpretation of PET data. Model-based interpretation of dynamic PET images by means of parametric fitting, however, is often a challenging task due to high levels of noise, thus necessitating a denoising step. The objective of this paper is to develop and characterize a denoising framework for dynamic PET based on non-local means (NLM). Theory NLM denoising computes weighted averages of voxel intensities assigning larger weights to voxels that are similar to a given voxel in terms of their local neighborhoods or patches. We introduce three key modifications to tailor the original NLM framework to dynamic PET. Firstly, we derive similarities from less noisy later time points in a typical PET acquisition to denoise the entire time series. Secondly, we use spatiotemporal patches for robust similarity computation. Finally, we use a spatially varying smoothing parameter based on a local variance approximation over each spatiotemporal patch. Methods To assess the performance of our denoising technique, we performed a realistic simulation on a dynamic digital phantom based on the Digimouse atlas. For experimental validation, we denoised PET images from a mouse study and a hepatocellular carcinoma patient study. We compared the performance of NLM denoising with four other denoising approaches–Gaussian filtering, PCA, HYPR, and conventional NLM based on spatial patches. Results The simulation study revealed significant improvement in bias-variance …