Diffusion Imaging in Python (Dipy) is a free and open source software projectfor the analysis of data from diffusion magnetic resonance imaging (dMRI)experiments. dMRI is an application of MRI that can be used to measurestructural features of brain white matter. Many methods have been developed touse dMRI data to model the local configuration of white matter nerve fiberbundles and infer the trajectory of bundles connecting different parts of thebrain.Dipy gathers implementations of many different methods in dMRI, including:diffusion signal pre-processing; reconstruction of diffusion distributions inindividual voxels; fiber tractography and fiber track post-processing, analysisand visualization. Dipy aims to provide transparent implementations forall the different steps of dMRI analysis with a uniform programming interface.We have implemented classical signal reconstruction techniques, such as thediffusion tensor model and deterministic fiber tractography. In addition,cutting edge novel reconstruction techniques are implemented, such asconstrained spherical deconvolution and diffusion spectrum imaging withdeconvolution, as well as methods for probabilistic tracking and originalmethods for tractography clustering. Many additional utility functions areprovided to calculate various statistics, informative visualizations, as wellas file-handling routines to assist in the development and use of noveltechniques.In contrast to many other scientific software projects, Dipy is not beingdeveloped by a single research group. Rather, it is an open project thatencourages contributions from any scientist/developer through GitHub and opendiscussions on the project mailing list. Consequently, Dipy today has aninternational team of contributors, spanning seven different academic institutionsin five countries and three continents, which is still growing.

Tractography based on non-invasive diffusion imaging is central to the study of human brain connectivity. To date, the approach has not been systematically validated in ground truth studies. Based on a simulated human brain data set with ground truth tracts, we organized an open international tractography challenge, which resulted in 96 distinct submissions from 20 research groups. Here, we report the encouraging finding that most state-of-the-art algorithms produce tractograms containing 90% of the ground truth bundles (to at least some extent). However, the same tractograms contain many more invalid than valid bundles, and half of these invalid bundles occur systematically across research groups. Taken together, our results demonstrate and confirm fundamental ambiguities inherent in tract reconstruction based on orientation information alone, which need to be considered when interpreting tractography and connectivity results. Our approach provides a novel framework for estimating reliability of tractography and encourages innovation to address its current limitations.

The ability of fiber tractography to delineate non‐invasively the white matter fiber pathways of the brain raises possibilities for clinical applications and offers enormous potential for neuroscience. In the last decade, fiber tracking has become the method of choice to investigate quantitative MRI parameters in specific bundles of white matter. For neurosurgeons, it is quickly becoming an invaluable tool for the planning of surgery, allowing for visualization and localization of important white matter pathways before and even during surgery. Fiber tracking has also claimed a central role in the field of “connectomics,” a technique that builds and studies comprehensive maps of the complex network of connections within the brain, and to which significant resources have been allocated worldwide. Despite its unique abilities and exciting applications, fiber tracking is not without controversy, in particular when it comes to its interpretation. As neuroscientists are eager to study the brain's connectivity, the quantification of tractography‐derived “connection strengths” between distant brain regions is becoming increasingly popular. However, this practice is often frowned upon by fiber‐tracking experts. In light of this controversy, this paper provides an overview of the key concepts of tractography, the technical considerations at play, and the different types of tractography algorithm, as well as the common misconceptions and mistakes that surround them. We also highlight the ongoing challenges related to fiber tracking. While recent methodological developments have vastly increased the biological accuracy of fiber tractograms, one should be aware that, even with state‐of‐the‐art techniques, many issues that severely bias the resulting structural “connectomes” remain unresolved.

Diffusion MRI‐based tractography is the most commonly‐used technique when inferring the structural brain connectome, i.e., the comprehensive map of the connections in the brain. The utility of graph theory—a powerful mathematical approach for modeling complex network systems—for analyzing tractography‐based connectomes brings important opportunities to interrogate connectome data, providing novel insights into the connectivity patterns and topological characteristics of brain structural networks. When applying this framework, however, there are challenges, particularly regarding methodological and biological plausibility. This article describes the challenges surrounding quantitative tractography and potential solutions. In addition, challenges related to the calculation of global network metrics based on graph theory are discussed.Evidence Level: 5Technical Efficacy: Stage 1

The human brain is a complex and organized network, where the connection between regions is not achieved with single axons crisscrossing each other but rather millions of densely packed and well-ordered axons. Reconstruction from diffusion MRI tractography is only an attempt to capture the full complexity of this network, at the macroscale. This review provides an overview of the misconceptions, biases and pitfalls present in structural white matter bundle and connectome reconstruction using tractography. The goal is not to discourage readers, but rather to inform them of the limitations present in the methods used by researchers in the field in order to focus on what they can do and promote proper interpretations of their results. It also provides a list of open problems that could be solved in future research projects for the next generation of PhD students.