Why has diffusion MRI become a principal modality for mapping connectomes in vivo? How do different image acquisition parameters, fiber tracking algorithms and other methodological choices affect connectome estimation? What are the main factors that dictate the success and failure of connectome reconstruction? These are some of the key questions that we aim to address in this review. We provide an overview of the key methods that can be used to estimate the nodes and edges of macroscale connectomes, and we discuss open problems and inherent limitations. We argue that diffusion MRI-based connectome mapping methods are still in their infancy and caution against blind application of deep white matter tractography due to the challenges inherent to connectome reconstruction. We review a number of studies that provide evidence of useful microstructural and network properties that can be extracted in various independent and biologically relevant contexts. Finally, we highlight some of the key deficiencies of current macroscale connectome mapping methodologies and motivate future developments. Functional integration, the interaction and information transfer between different subunits in the brain, is mediated in part through white matter connections. 1 The formation of these fiber pathways is guided by genetic, but also environmental, factors. During the early phases of development, an initial over-production of synapses is followed by pruning of the redundant connections in response to first life experiences. 2 The continuous maturation and myelination of white matter from the first months of life and through to adulthood reflects learning and interactions with external stimuli.This experience-dependent molding of brain connectivity 3 sheds light on the functional relevance of white matter pathways. Anatomical connections constrain neural computations. In fact, the pattern of anatomical connections a brain region has with other regions can predict, to a certain extent, the function of that region at a systems level. 4,5 This notion of connectivity fingerprinting and its functional implications has increased interest in studying connections and structural organization. 6 The term connectome, proposed roughly 10 years ago, 7,8 describes a comprehensive network map of extrinsic connections between functionally specialized brain regions. Ideally, such a map contains not only a list of connected areas, but also the relative strength and directionality of each connection. 8 Connectomics has the potential to reveal new insights into the principles that guide how different functional subunits are arranged and influence one another, 9 as well as how these processes are perturbed in pathological brain conditions. 10 Invasive approaches to map brain connections have existed for many decades. 11 At the microscale, techniques such as automated histological staining, 12,13 serial electron microscopy 14 and 3D fluorescence imaging 15 allow more data to be collected and processed nowadays with less laborintensive methods and fewer ima...