Increasing demand for quantitative diffusion MRI in biomedical research necessitates a reevaluation of potential sources of artifact. While cardiac pulsation-induced effects have been reported previously, their quantitative characterization under standard acquisition protocols remains outstanding. To determine whether practical data quality improvements merit the use of cardiac gating, the maximum effects induced in apparent diffusion coefficient, fractional anisotropy, and mean diffusivity measurements were quantitatively estimated via measurements and bootstrapping simulations. In apparent diffusion coefficient time courses, cardiac pulsation effects were measured to occupy approximately 6% of the heartbeat duration, with considerable intersession differences in effect magnitude. Bootstrapping revealed overestimations in fractional anisotropy up to 0.26, and mean diffusivity up to 0. The increasing demand for quantitative diffusion MRI in biomedical research and its potential application in clinical practice have highlighted the need to determine and optimize the precision and accuracy of measurements.Diffusion-weighted imaging (DWI) is a motion-sensitized variant of MRI, quantifying molecular Brownian motion along a single direction. In diffusion tensor imaging (DTI), a minimum of six non-collinear DWI measurements are used to characterize three-dimensional profiles of Brownian motion. These are commonly summarized as biologically relevant quantitative diffusion tensor (DT) indices, such as fractional anisotropy (FA) and mean diffusivity (MD) (1).The inherently high motion sensitivity of DWI makes measurements potentially susceptible to contamination by non-Brownian motion. While Brownian displacements during diffusion-encoding times of 50 ms (typical for contemporary human DWI scans) are approximately 10 lm (1,2), cardiac pulsation-induced displacements of up to 130 lm to 184 lm (3,4), with velocities of up to 1.5 mm sec À1 (3) have been reported. Single-direction DWI, scalar DT indices, and tractography results measured with single-shot echo-planar imaging sequences have been observed to exhibit cardiac pulsation-induced effects (5-12). Restricting DWI acquisitions to periods of reduced cardiac pulsation-induced motion, henceforth referred to as the ''still phase'', was found to prevent the occurrence of such effects (5-10,12).In numerous investigations (5,8-10,12,13), the number of measurable repeats was boosted by minimizing the number of DWI measurements per dataset. However, the biological relevance of single-direction DWI measurements is limited, and DTs are usually based on measurements along more than the minimum six directions (14,15). Therefore, while this approach is effective at demonstrating the susceptibility of DWI and DTI measures to cardiac pulsation, and the spatial distribution of induced errors, it remains qualitative. In a pilot investigation (7), cardiac pulsation was shown to induce notable errors in DT indices measured using a standard protocol with 15 gradient directions. However, a ...