This study highlights that organs can have variable sub-proteomes that do not necessarily correspond to anatomical features. The bovine heart as a model demonstrates at least 3-5 separate proteomes exist within a 2D plane of the heart. The main differences between these proteomes were the biochemical pathways utilized energy substrates. By mapping individual protein expression across a 2D image we can see where proteins of interest such as disease associated proteins are specifically expressed and how specific areas of the heart have subtle but significant differences in the ratio of mitochondrial complexes. This may provide an idea of where to focus analysis on in the cardiac proteome for future studies and into the different mechanisms associated with function, stresses and potential disease processes We set out to fully proteomically characterise a whole section of heart to see if the degree of proteome changes with very different anatomical and physiological function and which biochemical pathways are more prevalent or important in different areas and whether this could be explained by the physiological requirement of that area of the heart.