A real-time method of imaging glucose uptake in single, living mammalian cells
A real-time method of imaging glucose uptake in single, living mammalian cells
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DOI：
10.1038/nprot.2007.76
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发表时间：
2007-01-01
期刊：
影响因子：
14.8
通讯作者：
Inagaki, Nobuya
中科院分区：
文献类型：
--
作者：
Yamada, Katsuya;Saito, Mikako;Inagaki, Nobuya
该方案详细描述了使用荧光D-葡萄糖衍生物2-[N-（7-硝基苯-2-氧杂-1，3-二唑-4-基）氨基]-2-脱氧-D-葡萄糖（2-NBDG）作为示踪剂监测单个活哺乳动物细胞中葡萄糖摄取的方法。专门设计的室和灌流系统，用于评估2- NBDG摄取到细胞中的真实的时间，可以与其他荧光方法，如钙离子成像和随后的免疫荧光分类的细胞表现出分歧2- NBDG摄取。整个方案，包括免疫细胞化学，可在2 d内完成（细胞培养除外）。介绍了2-NBDG的合成方法。
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.