Related Experiment Videos
Bioimaging of nitric oxide with fluorescent indicators based on the rhodamine chromophore
H Kojima1, M Hirotani, N Nakatsubo
1Graduate School of Pharmaceutical Sciences, and Second Department of Internal Medicine, Faculty of Medicine, University of Tokyo, Japan.
Abstract:
Diaminofluoresceins are widely used for detection and imaging of nitric oxide (NO), but for biological applications, they have the disadvantages that the fluorescence of the fluorescein chromophore is pH-sensitive and overlaps the autofluorescence of cells. We have developed a membrane-permeable fluorescent indicator for NO based on the rhodamine chromophore, DAR-4M AM, which can be excited with 550-nm light. The fluorescence quantum yield of the product after reaction with NO is 840 times higher than that of DAR-4M. The detection limit of NO was 7 nM, and the fluorescence showed no pH dependency above pH 4. DAR-4M AM was successfully applied to practical bioimaging of NO produced in bovine aortic endothelial cells.