Protocol for determining glucose uptake and retention in mammalian cells

Glucose is central to metabolism and the principal energy source for the brain and red blood cells, and many tumors take up glucose avidly. Here, we present a protocol for determining glucose uptake using 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino)-2-deoxyglucose (2-NBDG), confocal imaging, and analysis in ImageJ. We describe steps for cell culture, treatment with 2-NBDG, and image acquisition. We then detail procedures for analyzing cellular 2-NBDG levels, providing an indicator for glucose uptake and retention in mammalian cells without using radioactive materials. For complete details on the use and execution of this protocol, please refer to He et al.¹.