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A New Highly Selective and Sensitive Assay for Fluorescence Imaging of . OH in Living Cells: Effectively Avoiding the Interference of Peroxynitrite
Author(s) -
Li Ping,
Xie Ting,
Duan Xia,
Yu Fabiao,
Wang Xu,
Tang Bo
Publication year - 2010
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.200901514
Subject(s) - peroxynitrite , fluorescence , chemistry , intracellular , reactive oxygen species , biophysics , radical , fluorescence lifetime imaging microscopy , membrane , cytotoxicity , photochemistry , biochemistry , in vitro , superoxide , biology , physics , quantum mechanics , enzyme
A new nonredox fluorescent probe to realize the imaging of hydroxyl radicals ( . OH) in living cells was designed and synthesized. The structure comprised the fluorescent dye boron dipyrromethene (BDP) and a 2,2,6,6‐tetramethyl‐1‐piperidinoxyl (TEMPO) unit. This probe could rapidly respond to . OH with a detection limit of 18 p M , and it possessed superior photostability and pH insensitivity. Other reactive oxygen species (ROS) and relevant intracellular components did not interfere. In particular, the important problem of ONOO − interference was efficiently avoided. An MTT assay proved that the probe was not very cytotoxic. The probe could penetrate into intact cell membranes to selectively detect intracellular . OH without causing cellular damage in living mice macrophages, normal human liver cells. and human hepatoma cells. These advantageous characteristics make the fluorescent probe potentially useful as a new candidate to detect . OH in broad biosystems.