Objective        To assess the possibility of a deoxyglucose analog 2-[N-(7-nitrobenz-2-oxa-1

3-diazol-4-yl) amino]-2-deoxyglucose (2-NBDG) as a fluorescence probe to detect the glucose uptake in H9c2 cells.       Methods       Glucose transporter 1 (GLUT1) and glucose transporter 4 (GLUT 4) proteins in H9c2 cells were identified by indirect immunofluorescence (IIF) assays. H9c2 cells were incubated with 2-NBDG at a cohort concentration for different periods and the most suitable concentration and exposure period were determined by fluorescence microplate reader. H9c2 cells were incubated with 100 μmol/L 2-NBDG for 30 min with or without D-Glu

a potent competitive inhibitor of 2-NBDG. After the incubation

the 2-NBDG uptake was observed and recorded by confocal laser scanning microscopy (CLSM) and flow cytometry (FCM).        Results       Over-expressed GLUT1 and GLUT 4 proteins were discovered in H9c2 cells. The most suitable concentration and exposure period for H9c2 cells incubation were 100 μmol/L 2-NBDG and 30 min. After incubated with 2-NBDG with or without D-Glu

bright green fluorescence of the 2-NBDG in H9c2 cells was observed under CLSM. The intracellular accumulation of 2-NBDG fluorescence was decreased to 27% (CLSM) and 46.7% (FCM) (P0.01) in groups of co-incubating with D-Glu. 2-NBDG was used to detect cell glucose uptake and confirmed that insulin to promote glucose uptake in H9c2 cells.       Conclusion        2-NBDG is quickly transported by GLUT1 and GLUT4 and accumulated in the H9c2 cells

which can be inhibited by D-Glu. 2-NBDG may be used as an optical probe for glucose uptake in H9c2 cells.