A method for fully automated morphological and topological quantification of microvascular structures in confocal laser scanning microscopy (CLSM) volume datasets is presented. Several characteristic morphological and topological quantities are calculated in a series of image-processing steps and can be used to compare single components as well as whole networks of microvascular structures to each other. The effect of the individual image-processing steps is illustrated and characteristic quantities of measured volume datasets are presented and discussed.