A real-time method of imaging glucose uptake in single, living mammalian cells
- 29 March 2007
- journal article
- Published by Springer Nature in Nature Protocols
- Vol. 2 (3) , 753-762
- https://doi.org/10.1038/nprot.2007.76
Abstract
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.Keywords
This publication has 23 references indexed in Scilit:
- Viable Cell Detection by the Combined Use of Fluorescent Glucose and Fluorescent GlycineBioscience, Biotechnology, and Biochemistry, 2003
- Confocal microscopy study of the different patterns of 2-NBDG uptake in rabbit enterocytes in the apical and basal zonePflügers Archiv - European Journal of Physiology, 2001
- Measurement of Glucose Uptake and Intracellular Calcium Concentration in Single, Living Pancreatic β-CellsJournal of Biological Chemistry, 2000
- A novel fluorescent derivative of glucose applicable to the assessment of glucose uptake activity of Escherichia coliBiochimica et Biophysica Acta (BBA) - General Subjects, 1996
- Differences in Glucose Transporter Gene Expression between Rat Pancreatic α- and β-Cells Are Correlated to Differences in Glucose Transport but Not in Glucose UtilizationJournal of Biological Chemistry, 1995
- The Use of Spectral Analysis to Determine Regional Cerebral Glucose Utilization with Positron Emission Tomography and [18F]Fluorodeoxyglucose: Theory, Implementation, and Optimization ProceduresJournal of Cerebral Blood Flow & Metabolism, 1994
- Sites and Mechanisms of Function-Related Changes in Energy Metabolism in the Nervous SystemDevelopmental Neuroscience, 1993
- Establishment of a Pancreatic β Cell Line That Retains Glucose-Inducible Insulin Secretion: Special Reference to Expression of Glucose Transporter Isoforms*Endocrinology, 1990
- Unique cytochalasin B binding characteristics of the hepatic glucose carrierBiochemistry, 1983
- THE [14C]DEOXYGLUCOSE METHOD FOR THE MEASUREMENT OF LOCAL CEREBRAL GLUCOSE UTILIZATION: THEORY, PROCEDURE, AND NORMAL VALUES IN THE CONSCIOUS AND ANESTHETIZED ALBINO RAT1Journal of Neurochemistry, 1977