Using Ultrafast Responsive Phosphorescent Nanoprobe to Visualize Elevated Peroxynitrite In Vitro and In Vivo via Ratiometric and Time-Resolved Photoluminescence Imaging

Peroxynitrite (ONOO^-), a potent biological oxidant, which has a short half-life in physiological conditions, is related to many diseases. Accurate peroxynitrite determination with superior selectivity and sensitivity is important for understanding biological roles of peroxynitrite in different health and disease tissues. Autofluorescence is an inevitable interference in luminescence biodetection and bioimaging, which often reduces signal-to-noise ratio during detection. In this work, a phosphorescent peroxynitrite nanoprobe (MSN-ONOO) which displays two emission bands is prepared by immobilizing two long-lived phosphorescent iridium(III) complexes that are peroxynitrite-activable and -inert, respectively, into water-dispersible mesoporous silica nanoparticles. Owing to the fast response rate, excellent sensitivity and outstanding selectivity of the nanoprobe toward peroxynitrite, it is further used for peroxynitrite determination in vitro and in vivo via ratiometric photoluminescence imaging. More notably, taking advantage of the long-lived phosphorescence of MSN-ONOO, in vivo elevated peroxynitrite is imaged with diminished autofluorescence interference and improved signal-to-noise ratio via time-resolved photoluminescence imaging. As far as it is known, this is the first time for endogenous peroxynitrite detection in vivo via the time-resolved photoluminescence imaging. Furthermore, the production of peroxynitrite in inflamed tissues is visualized.