Many diseases involve either the formation of new blood vessels (e.g., tumor angiogenesis) or the damage of existing ones (e.g., diabetic retinopathy) at the microcirculation level. Optical-resolution photoacoustic microscopy (OR-PAM), capable of imaging microvessels in 3D in vivo down to individual capillaries using endogenous contrast, has the potential to reveal microvascular information critical to the diagnosis and staging of microcirculation-related diseases. In this study, we have developed a dedicated microvascular quantification (MQ) algorithm for OR-PAM to automatically quantify multiple microvascular morphological parameters in parallel, including the vessel diameter distribution, the microvessel density, the vascular tortuosity, and the fractal dimension. The algorithm has been tested on in vivo OR-PAM images of a healthy mouse, demonstrating high accuracy for microvascular segmentation and quantification. The developed MQ algorithm for OR-PAM may greatly facilitate quantitative imaging of tumor angiogenesis and many other microcirculation related diseases in vivo. M. Arbeit, "VEGF is essential for hypoxia-inducible factor-mediated neovascularization but dispensable for endothelial sprouting," Proc. Natl. Acad. Sci. U. S. A. 108(32), 13264-13269 (2011). 9. J. Folkman, "Angiogenesis in cancer, vascular, rheumatoid and other disease," Nat. Med. 1(1), 27-30 (1995 1182-1186 (1971). 11. J. W. Baish and R. K. Jain, "Cancer, angiogenesis and fractals," Nat. Med. 4(9), 984 (1998). 12. R. K. Jain, "Normalizing tumor vasculature with anti-angiogenic therapy: A new paradigm for combination therapy," Nat. Med. 7(9), 987-989 (2001)