A real-time method of imaging glucose uptake in single, living mammalian cells

Nat Protoc. 2007;2(3):753-62. doi: 10.1038/nprot.2007.76.


This protocol details a method for monitoring glucose uptake into single, living mammalian cells using a fluorescent D-glucose derivative, 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose (2-NBDG), as a tracer. The specifically designed chamber and superfusion system for evaluating 2-NBDG uptake into cells in real time can be combined with other fluorescent methods such as Ca2+ imaging and the subsequent immunofluorescent classification of cells exhibiting divergent 2-NBDG uptake. The whole protocol, including immunocytochemistry, can be completed within 2 d (except for cell culture). The procedure for 2-NBDG synthesis is also presented.

MeSH terms

  • 4-Chloro-7-nitrobenzofurazan / analogs & derivatives*
  • 4-Chloro-7-nitrobenzofurazan / chemical synthesis
  • 4-Chloro-7-nitrobenzofurazan / metabolism
  • Animals
  • Biological Transport / physiology
  • Cell Line
  • Deoxyglucose / analogs & derivatives*
  • Deoxyglucose / chemical synthesis
  • Deoxyglucose / metabolism
  • Glucose / metabolism*
  • Immunohistochemistry
  • Kinetics
  • Mice
  • Microscopy, Fluorescence


  • Deoxyglucose
  • 4-Chloro-7-nitrobenzofurazan
  • Glucose
  • 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose