Sensitivity evaluation of rhodamine B hydrazide towards nitric oxide and its application for macrophage cells imaging

  • Anal Chim Acta. 2011 Dec 5;708(1-2):141-8. doi: 10.1016/j.aca.2011.10.005.
Chi-Ming Wu  1 ,  Yen-Hao Chen ,  Kasala Dayananda ,  Tsun-Wei Shiue ,  Chen-Hsiung Hung ,  Wen-Feng Liaw ,  Po-Yu Chen ,  Yun-Ming Wang
Affiliations
  • 1. Department of Medicinal and Applied Chemisrty, Kaohsiung Medical University, Kaohsiung 807, Taiwan.
PMID: 22093357 DOI: 10.1016/j.aca.2011.10.005
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.

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