title: Sensitivity evaluation of rhodamine B hydrazide towards nitric oxide and its application for macrophage cells imaging.
abstract: A colorless and non-fluorescent rhodamine derivative, rhodamine B hydrazide (RH), is applied to detect nitric oxide and form fluorescent rhodamine B (RB). The reaction mechanism of RH with NO is proposed in this study. The probe shows good stability over a broad pH range (pH>4). Furthermore, fluorescence intensity of RH displays an excellent linearity to the NO concentration and the detection limit is as low as 20 nM. A 1000-fold fluorescence turn-on from a dark background was observed. Moreover, the selectivity study indicated that the fluorescence intensity increasing in the presence of NO was significantly higher than those of other reactive oxygen/nitrogen species. In exogenously generated NO detection study, clear intracellular red fluorescence was observed in the presence of S-nitroso-N-acetyl-D,L-penicillamine (SNAP, a kind of NO releasing agent). In endogenously generated NO detection study, increasing incubation time of RH with lipopolysaccharied (LPS) pre-treated cells could obtain a highly fluorescent cell image. These cell imaging results demonstrated that RH can efficiently penetrate into Raw 264.7 cells and be used for detection of exogenously and endogenously generated nitric oxide.
authors.affiliation: Department of Medicinal and Applied Chemisrty, Kaohsiung Medical University, Kaohsiung 807, Taiwan.
authors.forename: Chi-Ming
authors.initials: CM
authors.lastname: Wu
authors.forename: Yen-Hao
authors.initials: YH
authors.lastname: Chen
authors.forename: Kasala
authors.initials: K
authors.lastname: Dayananda
authors.forename: Tsun-Wei
authors.initials: TW
authors.lastname: Shiue
authors.forename: Chen-Hsiung
authors.initials: CH
authors.lastname: Hung
authors.forename: Wen-Feng
authors.initials: WF
authors.lastname: Liaw
authors.forename: Po-Yu
authors.initials: PY
authors.lastname: Chen
authors.forename: Yun-Ming
authors.initials: YM
authors.lastname: Wang
mesh_terms: D000818:Animals; D002460:Cell Line; D006834:Hydrazines / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008070:Lipopolysaccharides / Q000633:toxicity; D008264:Macrophages / Q000166:cytology*; D051379:Mice; D008856:Microscopy, Fluorescence; D009569:Nitric Oxide / Q000378:metabolism*; D012235:Rhodamines / Q000737:chemistry*; D026423:S-Nitroso-N-Acetylpenicillamine / Q000737:chemistry; D013997:Time Factors
chemical_list: D006834:Hydrazines; D008070:Lipopolysaccharides; D012235:Rhodamines; C543681:rhodamine B hydrazide; D009569:Nitric Oxide; D026423:S-Nitroso-N-Acetylpenicillamine