Two-photon phosphorescence lifetime microscopy allows depth-resolved micron-scale measurements of oxygen partial pressure (Po2) in the brain. The spatiotemporal resolution of these measurements has shown that the portion of capillary plasma in the vicinity of red blood cells (RBCs) has a higher Po2(Po2RBC) than that distant from RBCs(Po2InterRBC). Our group has shown that Po2InterRBC equilibrates with neuropil Po2 and can thus be used to non-invasively measure tissue Po2 (Parpaleix et al., 2013). The relevance of reported high-resolution Po2 values remains uncertain as measurements have only been performed during anaesthesia, which affects both neuronal activity and cerebral blood flow, and thus brain Po2.I measured Po2 at rest, in the awake, unstressed mouse in two brain regions, the olfactory bulb glomerular layer (GL) and the somatosensory cortex. The first section of my research, conducted in the GL, produced the first measurements of blood flow and Po2 parameters in the cerebral microvasculature in physiological conditions. I determined mean Po2 levels of values of 60.6 mmHg for Po2RBC, 23 mmHg for tissue Po2. In the cortex Po2 values show differences between the superficial layers. Furthermore the relationships of RBC Po2 and tissue Po2 to the blood flow parameters differed between the cortical layers, and also in comparison to the olfactory bulb GL.I compared both vascular and tissue Po2 between the awake and anaesthetised states, and observed that anaesthetics can dramatically change Po2 at the microvascular scale. This finding emphasises the importance of measuring these values in the physiologically normal brain.