Blood flow and its payload of molecular oxygen are two parameters of most physiological interest. Systemic tissue health is routinely gauged through measurements of vitals and oxygen saturation to estimate the state of these physiological parameters. We design, develop, and deploy optical imaging systems for examining perfusion and oxygenation at the local tissue level and apply these techniques for elucidating the normal and pathological processes associated with neurovascular disease. Specifically, we develop and validate the ability to use Multi-Exposure Speckle Imaging (MESI) to estimate microvascular flow dynamics in rodents over acute and chronic periods. Next, we pose significant optimizations to improve the efficacy of the widefield imaging technique for adoption by bench-side and clinical perfusion studies. We also introduce re-interpretations of the underlying physics to advance the theory that quantifies motion from the imaged speckle patterns. Finally, the technique is deployed for chronic monitoring of cortical flow dynamics before after focal ischemia of the motor cortex as part of a behavioral study in rodents. At the microscale, we develop and validate Two Photon Phosphorescence Lifetime Microscopy (2PLM) to examine dissolved oxygen concentration in microvasculature in three dimensions. We examine the technique’s ability for functional mapping of the rodent cortical microvascular network by quantifying the partial pressure of oxygen (pO₂) before and after occlusion of critical arterioles. Automation of acquisitions and processing for robust oxygen mapping within the micro-vascular network are developed and evaluated. The in vivo results are presented in light of those from studies utilizing more invasive mapping electrodes to provide independent corroboration of the observed neurovascular oxygen distributions. The technique is deployed for examining high resolution functional and structural remodeling after focal cerebral ischemia. / text