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Proceedings Paper

In vivo quantitative visualization of hypochlorous acid in the liver using a novel selective two-photon fluorescent probe
Author(s): Haolu Wang; Aparna Jayachandran; Germain Gravot; Xiaowen Liang; Camilla A. Thorling; Run Zhang; Xin Liu; Michael S. Roberts
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Paper Abstract

Hypochlorous acid (HOCl) plays a vital role in physiological events and diseases. During hepatic ischemia-reperfusion (I/R) injury, HOCl is generated by neutrophils and diffuses into hepatocytes, causing oxidant stress-mediated injury. Although many probes have been developed to detect HOCl, most were difficult to be distinguished from endogenous fluorophores in intravital imaging and only can be employed under one-photon microscopy. A novel iridium(III) complex-based ferrocene dual-signaling chemosensor (Ir-Fc) was designed and synthesized. Ir-Fc exhibited a strong positive fluorescent response only in the presence of HOCl, whereas negligible fluorescent signals were observed upon the additions of other reactive oxygen/nitrogen species and metal ions. There was a good linear relationship between probe responsive fluorescent intensity and HOCl concentration. Ir-Fc was then intravenously injected into BALB/c mice at the final concentration of 50 μM and the mouse livers were imaged using multiphoton microscopy (MPM). In the I/R liver, reduced autofluorescence was detected by MPM, indicating the hepatocyte necrosis. Remarkable enhancement of red fluorescence was observed in hepatocytes with decreased autofluorescence, indicating the reaction of Ir-Fc with endogenous HOCl molecules. The cellular concentration of HOCl was first calculated based on the intensity of MPM images. No obvious toxic effects were observed in histological examination of major organs after Ir-Fc injection. In summary, Ir-Fc has low cytotoxicity, high specificity to HOCl, and rapid “off-on” fluorescence. It is suitable for dynamic quantitatively monitoring HOCl generation using MPM at the cellular level. This technique can be readily extended to examination of liver diseases and injury.

Paper Details

Date Published: 24 November 2016
PDF: 3 pages
Proc. SPIE 10013, SPIE BioPhotonics Australasia, 100131G (24 November 2016); doi: 10.1117/12.2242978
Show Author Affiliations
Haolu Wang, The Univ. of Queensland (Australia)
Aparna Jayachandran, The Univ. of Queensland (Australia)
Germain Gravot, Univ. de Rennes 1 (France)
Xiaowen Liang, The Univ. of Queensland (Australia)
Camilla A. Thorling, The Univ. of Queensland (Australia)
Run Zhang, The Univ. of Queensland (Australia)
Xin Liu, The Univ. of Queensland (Australia)
Michael S. Roberts, The Univ. of Queensland (Australia)
The Univ. of South Australia (Australia)

Published in SPIE Proceedings Vol. 10013:
SPIE BioPhotonics Australasia
Mark R. Hutchinson; Ewa M. Goldys, Editor(s)

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