The utility of single-pulse electrical stimulation (SPES) for epilepsy surgery has been highlighted in the last decade. When applied at a frequency of about 1 Hz, it can probe cortico-cortical connections by averaging electrocorticographic signal time-locked to stimuli to record cortico-cortical evoked potentials (CCEPs) emanating from adjacent and remote cortices. Although limited to patients undergoing invasive presurgical evaluations, CCEPs provide a novel way to explore inter-regional connectivity in vivo in the living human brain to probe functional brain networks such as language and cognitive motor networks. In addition to its impact on basic systems neuroscience, this method, in combination with 50 Hz electrical cortical stimulation, can contribute clinically to the mapping of functional brain systems by tracking cortico-cortical connections among functional cortical regions in individual patients. This approach may help identify normal cortico-cortical networks in pathological brain, or plasticity of brain systems in conjunction with pathology. Because of its high practical value, it has been applied to intraoperative monitoring of functional brain networks in patients with brain tumours. With regard to epilepsy, SPES has been used to probe cortical excitability of the focus (epileptogenicity) and seizure networks. Both early (i.e. CCEP) and delayed responses are regarded as surrogate markers of epileptogenicity. With regard to its potential impact on human brain connectivity maps, worldwide collaboration is warranted to establish standardized CCEP connectivity maps as a solid reference for non-invasive connectome research.