title: Bioimaging of nitric oxide with fluorescent indicators based on the rhodamine chromophore.
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.
authors.affiliation: Graduate School of Pharmaceutical Sciences, and Second Department of Internal Medicine, Faculty of Medicine, University of Tokyo, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Kojima
authors.forename: M
authors.initials: M
authors.lastname: Hirotani
authors.forename: N
authors.initials: N
authors.lastname: Nakatsubo
authors.forename: K
authors.initials: K
authors.lastname: Kikuchi
authors.forename: Y
authors.initials: Y
authors.lastname: Urano
authors.forename: T
authors.initials: T
authors.lastname: Higuchi
authors.forename: Y
authors.initials: Y
authors.lastname: Hirata
authors.forename: T
authors.initials: T
authors.lastname: Nagano
mesh_terms: D000818:Animals; D001011:Aorta / Q000166:cytology; D001921:Brain / Q000378:metabolism; D002417:Cattle; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D004730:Endothelium, Vascular / Q000737:chemistry / Q000166:cytology; D005456:Fluorescent Dyes / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D015394:Molecular Structure; D009569:Nitric Oxide / Q000032:analysis*; D010784:Photomicrography; D051381:Rats; D017207:Rats, Sprague-Dawley; D012235:Rhodamines / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D005456:Fluorescent Dyes; D012235:Rhodamines; D009569:Nitric Oxide