title: Real-Time Discrimination and Versatile Profiling of Spontaneous Reactive Oxygen Species in Living Organisms with a Single Fluorescent Probe.
abstract: Fluorescent probes are powerful tools for the investigations of reactive oxygen species (ROS) in living organisms by visualization and imaging. However, the multiparallel assays of several ROS with multiple probes are often limited by the available number of spectrally nonoverlapping chromophores together with large invasive effects and discrepant biological locations. Meanwhile, the spontaneous ROS profilings in various living organs/tissues are also limited by the penetration capability of probes across different biological barriers and the stability in reactive in vivo environments. Here, we report a single fluorescent probe to achieve the effective discrimination and profiling of hydroxyl radicals (•OH) and hypochlorous acid (HClO) in living organisms. The probe is constructed by chemically grafting an additional five-membered heterocyclic ring and a lateral triethylene glycol chain to a fluorescein mother, which does not only turn off the fluorescence of fluorescein, but also create the dual reactive sites to ROS and the penetration capability in passing through various biological barriers. The reactions of probe with •OH and HClO simultaneously result in cyan and green emissions, respectively, providing the real-time discrimination and quantitative analysis of the two ROS in cellular mitochondria. Surprisingly, the accumulation of probes in the intestine and liver of a normal-state zebrafish and the transfer pathway from intestine-to-blood-to-organ/tissue-to-kidney-to-excretion clearly present the profiling of spontaneous •OH and HClO in these metabolic organs. In particular, the stress generation of •OH at the fresh wound of zebrafish is successfully visualized for the first time, in spite of its extremely short lifetime.
authors.affiliation: CAS Center for Excellence in Nanoscience, Institute of Intelligent Machines, Chinese Academy of Sciences , Hefei, Anhui 230031, China.|Department of Chemistry, University of Science and Technology of China , Hefei, Anhui 230026, China.
authors.forename: Ruilong
authors.initials: R
authors.lastname: Zhang
authors.affiliation: CAS Center for Excellence in Nanoscience, Institute of Intelligent Machines, Chinese Academy of Sciences , Hefei, Anhui 230031, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Zhao
authors.affiliation: CAS Center for Excellence in Nanoscience, Institute of Intelligent Machines, Chinese Academy of Sciences , Hefei, Anhui 230031, China.
authors.forename: Guangmei
authors.initials: G
authors.lastname: Han
authors.affiliation: CAS Center for Excellence in Nanoscience, Institute of Intelligent Machines, Chinese Academy of Sciences , Hefei, Anhui 230031, China.|Department of Chemistry, University of Science and Technology of China , Hefei, Anhui 230026, China.
authors.forename: Zhengjie
authors.initials: Z
authors.lastname: Liu
authors.affiliation: CAS Center for Excellence in Nanoscience, Institute of Intelligent Machines, Chinese Academy of Sciences , Hefei, Anhui 230031, China.|Department of Chemistry, University of Science and Technology of China , Hefei, Anhui 230026, China.
authors.forename: Cui
authors.initials: C
authors.lastname: Liu
authors.affiliation: CAS Center for Excellence in Nanoscience, Institute of Intelligent Machines, Chinese Academy of Sciences , Hefei, Anhui 230031, China.|Department of Chemistry, University of Science and Technology of China , Hefei, Anhui 230026, China.
authors.forename: Cheng
authors.initials: C
authors.lastname: Zhang
authors.affiliation: CAS Center for Excellence in Nanoscience, Institute of Intelligent Machines, Chinese Academy of Sciences , Hefei, Anhui 230031, China.
authors.forename: Bianhua
authors.initials: B
authors.lastname: Liu
authors.affiliation: CAS Center for Excellence in Nanoscience, Institute of Intelligent Machines, Chinese Academy of Sciences , Hefei, Anhui 230031, China.
authors.forename: Changlong
authors.initials: C
authors.lastname: Jiang
authors.affiliation: CAS Center for Excellence in Nanoscience, Institute of Intelligent Machines, Chinese Academy of Sciences , Hefei, Anhui 230031, China.
authors.forename: Renyong
authors.initials: R
authors.lastname: Liu
authors.affiliation: CAS Center for Excellence in Nanoscience, Institute of Intelligent Machines, Chinese Academy of Sciences , Hefei, Anhui 230031, China.
authors.forename: Tingting
authors.initials: T
authors.lastname: Zhao
authors.affiliation: CAS Center for Excellence in Nanoscience, Institute of Intelligent Machines, Chinese Academy of Sciences , Hefei, Anhui 230031, China.|Institute of Materials Research and Engineering , A-STAR, 3 Research Link, Singapore 117602.
authors.forename: Ming-Yong
authors.initials: MY
authors.lastname: Han
authors.affiliation: CAS Center for Excellence in Nanoscience, Institute of Intelligent Machines, Chinese Academy of Sciences , Hefei, Anhui 230031, China.|Department of Chemistry, University of Science and Technology of China , Hefei, Anhui 230026, China.|State Key Laboratory of Transducer Technology, Chinese Academy of Sciences , Hefei, Anhui 230031, China.
authors.forename: Zhongping
authors.initials: Z
authors.lastname: Zhang
mesh_terms: D000818:Animals; D003199:Computer Systems; D005452:Fluoresceins / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D017665:Hydroxyl Radical / Q000032:analysis / Q000378:metabolism; D006997:Hypochlorous Acid / Q000032:analysis / Q000378:metabolism; D051379:Mice; D011092:Polyethylene Glycols / Q000737:chemistry; D000067996:RAW 264.7 Cells; D017382:Reactive Oxygen Species / Q000032:analysis* / Q000378:metabolism; D013050:Spectrometry, Fluorescence / Q000379:methods; D014947:Wounds and Injuries / Q000378:metabolism; D015027:Zebrafish
chemical_list: D005452:Fluoresceins; D005456:Fluorescent Dyes; D017382:Reactive Oxygen Species; C052067:fluorescein hydrazide; D017665:Hydroxyl Radical; C028914:triethylene glycol; D011092:Polyethylene Glycols; D006997:Hypochlorous Acid