Purpose: We studied the topographic distribution of contractile protein in different orders of the human macular microvasculature to further understanding of the sites for capillary blood flow regulation. Methods: Nine donor eyes from eight donors were cannulated at the central retinal artery and perfusion labeled for alpha smooth muscle actin (αSMA) and filamentous actin (F-actin). Confocal images were collected from the macula region, viewed, projected, and converted to a 255 grayscale for measurements. The mean intensity was measured for macular arterioles, venules, and capillary segments. The diameter of each vessel segment measured was recorded. The normalized mean intensity values from all images were ranked according to vessel types and size with a total of nine categories. Results: F-actin was present throughout the macular microvasculature whereas αSMA labeling showed variations. Overall, αSMA has a more prominent presence in the macular arterioles than in the macular capillaries and venules, and αSMA strongly labeled the smaller macular arterioles. Some capillaries also labeled positive for αSMA, including some of the capillaries in the innermost capillary ring surrounding the foveola. It was weakly present in the capillaries on the venous side and larger venules. In the larger macular arterioles closer to 100 μm in diameter, αSMA labeling was weakly present and not as ubiquitous as in the smaller arterioles. Conclusions: Nonuniform distribution of contractile proteins in the different types, orders, and sizes of macular microvasculature indicates that these vessels may have different contractile capability and roles in macular flow regulation.