The 5’-AMP-activated protein kinase (AMPK) functions as an intracellular fuel sensor and is activated in skeletal muscle in response to exercise and utilization of stored energy. AMPK is a heterotrimer, consisting of a-, b-, and g-subunits. The AMPK b-subunit has been intricately linked to glycogen regulation in skeletal muscle due to its carbohydrate binding motif and evidence that AMPK binds glycogen in vitro and the fact that glycogen is used during exercise. To determine the physiological relevance of this, a single muscle fibre approach was adopted, and the movement of AMPK and glycogen assessed. In rodent skeletal muscle, it was found that there is no association of AMPK with glycogen, and this will be discussed in detail. Further exploration of this potential relationship in human skeletal muscle was undertaken, and of particular note the findings in individuals with type 2 diabetes (T2D) will be discussed. With those findings, overall, it is demonstrated that the regulation of glucose metabolism and glycogen storage is fibre-type specific in skeletal muscle, with significant differences between muscle from control and individuals with T2D.