As a preliminary to analysing substrate utilization by the rodent retina in various functional states, the in vivo uptake of 2-deoxy-D-glucose in light- and dark-adapted rat retinas was studied. This glucose analog can be phosphorylated, but not further metabolized, and the rate of its intracellular accumulation can be used as an index of glucose utilisation in the tissue. The results localize the decreased uptake of 2-deoxy glucose to photoreceptors and imply a decreased utilization of glucose when these cells are exposed to steady illumination.