Recently, the application of neuroscience methods and findings to the study of organizational phenomena has gained significant interest and converged in the emerging field of organizational neuroscience. Yet, this body of research has principally focused on the brain, often overlooking fuller analysis of the activities of the human nervous system and associated methods available to assess them. In this article, we aim to narrow this gap by reviewing heart rate variability (HRV) analysis, which is that set of methods assessing beat-to-beat changes in the heart rhythm over time, used to draw inference on the outflow of the autonomic nervous system (ANS). In addition to anatomo-physiological and detailed methodological considerations, we discuss related theoretical, ethical, and practical implications. Overall, we argue that this methodology offers the opportunity not only to inform on a wealth of constructs relevant for management inquiries but also to advance the overarching organizational neuroscience research agenda and its ecological validity.