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Synthesis of a highly HOCl-selective fluorescent probe and its use for imaging HOCl in cells and organisms.

Authors
 Xiaoqiang Chen  ;  Kyung-Ah Lee  ;  Xintong Ren  ;  Jae-Chan Ryu  ;  Gyungmi Kim  ;  Ji-Hwan Ryu  ;  Won-Jae Lee  ;  Juyoung Yoon 
Citation
 NATURE PROTOCOLS, Vol.11(7) : 1219-1228, 2016 
Journal Title
NATURE PROTOCOLS
ISSN
 1754-2189 
Issue Date
2016
MeSH
Animals ; Cells, Cultured ; Drosophila/chemistry ; Drosophila/microbiology ; Fluorescent Dyes/chemical synthesis ; Fluorescent Dyes/chemistry* ; Hypochlorous Acid/analysis* ; Intestines/chemistry ; Intestines/microbiology ; Mice ; Mice, Inbred C57BL ; Microscopy, Confocal/methods ; Neutrophils/chemistry ; Neutrophils/cytology ; Optical Imaging/methods* ; Rhodamines/chemical synthesis ; Rhodamines/chemistry*
Abstract
During infection, nicotinamide adenine dinucleotide phosphate-oxidase of innate immune cells generates important microbicidal reactive oxygen species (ROS) such as hypochlorous acid (HOCl) to kill the invading pathogens. However, excess amounts of HOCl induce oxidative damage of functional biomolecules such as DNA and proteins, which may cause chronic inflammatory diseases. Herein, we outline protocols for the preparation of a rhodamine-based HOCl probe, as well as applications thereof, with which to detect HOCl in living cells and organisms. The probe (R19S) can be prepared from a commercially available rhodamine, rhodamine 6G, in two steps. When R19S is treated with HOCl, the sulfur atom is replaced by an oxygen atom, resulting in opening of the lactone ring; thus, nonfluorescent R19S is converted to highly fluorescent rhodamine 19 (R19). R19S exhibits high selectivity for HOCl over other ROS and high sensitivity in a weakly acidic environment. In addition, we describe fluorescence imaging assays of HOCl in mouse neutrophils and Drosophila targeted using this probe. The approximate amount of time required to synthesize the probe is 2-3 d, after which it can be used for up to 5 h in the bioimaging of living cells.
Full Text
http://www.nature.com/nprot/journal/v11/n7/full/nprot.2016.062.html
DOI
10.1038/nprot.2016.062
Appears in Collections:
1. College of Medicine (의과대학) > BioMedical Science Institute (의생명과학부) > 1. Journal Papers
Yonsei Authors
Ryu, Ji Hwan(유지환)
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/147079
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