Macromolecule diffusion in cells and tissues is important for cell signaling, metabolism and locomotion. Biophysical methods, including non-invasive or minimally invasive in-vivo photobleaching techniques and single quantum-dot tracking, have been used to measure the rates of macromolecule diffusion in living cells and tissues, including central nervous system and tumors. Mathematical modeling and statistical analysis of experimental data revealed various modes of diffusion, which are strongly coupled with spatiotemporal changes in nanoscale structures and material properties.