The vasculature ensures optimal delivery of nutrients and oxygen throughout the body. The ability to respond to changing tissue demands requires constant reshaping of the vascular network through modulation of its density, diameter, or patterning. These processes are especially prominent after tissue damage or in tumors. The matrix metalloproteinase (MMP) family of endopeptidases are key contributors to vascular remodeling, able to cleave all extracellular matrix components and also soluble factors and membrane receptors. Observations recorded over several decades have established that the vasculature changes in pathological contexts, and this has formed the basis for developing angiotherapies as a novel approach to treating disease. For example, inhibition of angiogenesis or normalization of the vasculature has been proposed as treatment for cancer and chronic inflammatory diseases. In contrast, boosting angiogenesis may be helpful in ischemic conditions such as myocardial infarction and in regenerative medicine. Classical histological methods for the analysis of tissue vasculature have relied on thin sections that do not capture the complex 3D structure of the vascular network. Given the importance of understanding disease-associated vascular changes for the development of rational angiotherapeutic interventions, we present a protocol for thick section-based 3D image analysis of vasculature structure and function.
Keywords: Confocal microscope; Mouse; Quantification; Thick section; Three-dimensional; Vasculature; Volumetric.