Muscle fatigue has been studied with a variety approaches, tools and technologies. The foci of these studies have ranged tremendously, from molecules to the entire organism. Single cell and animal models have been used to gain mechanistic insight into the fatigue process. The theme of this review is the concept that the mechanisms of muscle fatigue do not occur in isolation in vivo: muscular work is supported by many complex physiological systems, any of which could fail during exercise and thus contribute to fatigue. To advance our overall understanding of fatigue, a combination of models and approaches is necessary. In this review, we examine the roles that neuromuscular properties, intracellular glycogen, oxygen metabolism, and blood flow play in the fatigue process during exercise and pathological conditions.