The microfluidic device can control both the spatial and temporal variations in oxygen tensions to better replicate in vivo biology. The device consists of three parallel connected tissue chambers and an oxygen scavenger channel placed adjacent to these tissue chambers. Experimentally measured oxygen maps were constructed using phosphorescent lifetime imaging microscopy and compared with values from a computational model. For testing, it was coupled with a 3D sprouting angiogenesis assay. The simple design provides consistent control of spatial and temporal oxygen gradients in the tissue microenvironment and can be used to investigate important oxygen-dependent biological processes.