16Diffusion MRI tractography is commonly used to delineate white matter tracts. These delineations can 17 be used for planning neurosurgery or for identifying regions of interest from which microstructural 18 measurements can be taken. Probabilistic tractography produces different delineations each time it 19 is run, potentially leading to microstructural measurements or anatomical delineations that are not 20 reproducible. Generating a sufficiently large number of streamlines is required to avoid this scenario, 21 but what constitutes "sufficient" is difficult to assess and so streamline counts are typically chosen in 22 an arbitrary or qualitative manner. This work explores several factors influencing tractography 23 reliability and details two methods for estimating this reliability. The first method automatically 24 estimates the number of streamlines required to achieve reliable microstructural measurements, 25 whilst the second estimates the number of streamlines required to achieve a reliable binarised 26 trackmap than can be used clinically. Using these methods, we calculated the number of streamlines 27 required to achieve a range of quantitative reproducibility criteria for three anatomical tracts in 40 28Human Connectome Project datasets. Actual reproducibility was checked by repeatedly generating 29 the tractograms with the calculated numbers of streamlines. We found that the required number of 30 streamlines varied strongly by anatomical tract, image resolution, number of diffusion directions, the 31 degree of reliability desired, the microstructural measurement of interest, and/or the specifics on how 32 the tractogram was converted to a binary volume. The proposed methods consistently predicted 33 streamline counts that achieved the target reproducibility. Implementations are made available to 34 enable the scientific community to more-easily achieve reproducible tractography. 35 Keywords 36 diffusion weighted imaging; diffusion tractography; power analysis; streamline count; bootstrapping; 37 reproducibility. 38 39 40 3