A real-time method of imaging glucose uptake in single, living mammalian cells
K Yamada, M Saito, H Matsuoka, N Inagaki - Nature protocols, 2007 - nature.com
K Yamada, M Saito, H Matsuoka, N Inagaki
Nature protocols, 2007•nature.comThis 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 …
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 …
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.
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