ESIPT-based ratiometric fluorescence probe for the intracellular imaging of peroxynitrite

Abstract

In this work, we set out to develop an endoplasmic reticulum (ER) directed ESIPT-based ONOO ratiometric fluorescent probe (ABAH-LW). ABAH-LW was synthesized in four steps and found to have a high sensitivity and selectivity towards the detection of ONOO. ABAH-LW was able to detect low concentrations of ONOO (limit of detection = 21.4 nM) within seconds producing a ratiometric change in fluorescence intensity. ABAH-LW further demonstrated the ability to ratiometrically image endogenous and exogenous ONOO in HeLa cells. Moreover, co-localization experiments were carried out using commercially available ER-Tracker Red, Lyso-Tracker Red and Mito-Tracker-Red, which were co-stained with ABAH-LW in HeLa cells. For ER-Tracker Red, Pearson's correlation co-efficient of 0.93 was determined and 3D surface plot analysis illustrated a large overlap between ABAH-LW and ER-Tracker Red using both red and blue channels. In addition some co-localisation with Mito-Tracker Red and ABAH-LW was observed (0.73).

Excited-state intramolecular proton-transfer (ESIPT) based fluorescence sensors and imaging agents

“…83 More recently, James and co-workers developed a ratiometric boronate-based ESIPT probe for the visualisation of ONOO À within the endoplasmic reticulum. 84 Sulfites (SO 3…”

ESIPT-based fluorescence probe for the ratiometric detection of superoxide

“…[7][8][9] Within our research group, we have developed several ESIPT-based fluorescent probes for the detection of biological reactive oxygen species as well as biological thiols. [10][11][12][13] Previously, we have developed a thiocarbamate functionalised methoxyhydroxybenzothiazole (HMBT) fluorescent probe TCBT-OMe for the detection of HOCl/ClO À (Scheme 1). The addition of HOCl/ ClO À to TCBT-OMe resulted in the rapid hydrolysis (o10 s) of the thiocarbamate linker, leading to a ratiometric change in fluorescence intensity.…”

Sensing Peroxynitrite in Different Organelles of Murine RAW264.7 Macrophages With Coumarin-Based Fluorescent Probes

“…The development of fluorescent tools to monitor reactive oxygen species (ROS), in particular ONOO − , has lead in recent years to the development of a variety of diagnostic small molecule fluorescent probes (Wu et al, 2017;Wang et al, 2018). Our group has exploited boronic acids and esters as a sensing motif for ONOO − (Odyniec et al, 2018;Weber et al, 2018;Wu et al, 2018). The fluorophore is masked by boronate esters which upon reaction with ONOO − results in fluorophore activation and the emission of a distinctive fluorescence signal (Scheme 1).…”