Synthesis of a highly metal-selective rhodamine-based probe and its use for the in vivo monitoring of mercury

Abstract

This protocol describes detailed procedures for the preparation of a rhodamine-based mercury probe and for its applications to the detection of mercury in cells and vertebrate organisms. The mercury probe 1, which is prepared in two steps from rhodamine 6G, responds rapidly to Hg 2+ in aqueous solutions with a 1:1 stoichiometry. Owing to the fact that the probe reacts with Hg 2+ in an irreversible manner, it has advantages over other reversible mercury probes in in vivo assays with respect to both sensitivity and selectivity. In addition, fluorescent imaging assays of Hg 2+ in live cells and zebrafish by using this mercury probe are detailed in this protocol. The approximate time frame for the preparation of the probe is 24 h and for its use in imaging assays is 1.5 h.

Highly Selective and Ultrasensitive Turn-on Luminescence Chemosensor for Mercury (II) Determination Based on the Rhodamine 6G Derivative FC1 and Au Nanoparticles

“…FC1 was synthesized following the procedure previously described in the literature [10,12,18]. The stock solution of FC1 1 mmol·L −1 was prepared by dissolving 5.0 mg of FC1 in 10.00 mL of ACN.…”

“…Rhodamine 6G spirocyclic phenylthiosemicarbazide derivative (FC1) has been previously proposed for Hg 2+ determination in diverse matrices, such as water and fish samples [10,11,12], reporting a limit of detection of 1 μg·L −1 [10]. Unlike other chemosensors, the probe responds to Hg 2+ in an irreversible manner, as the rhodamine 6G derivative undergoes oxadiazole formation when the thiosemicarbazide moiety is liberated by Hg 2+ , facilitating ring opening of the spirocycle group.…”

A cyanine-modified upconversion nanoprobe for NIR-excited imaging of endogenous hydrogen peroxide signaling in vivo

“…Almost all the eggs were fertilized immediately. The 72 hpf old zebrafish was maintained in E3 embryo media (15 mM NaCl, 0.5 mM KCl, 1 mM MgSO 4 , 1 mM CaCl 2 , 0.15 mM KH 2 PO 4 , 0.05 mM Na 2 HPO 4 , 0.7 mM NaHCO 3 , 10-5% methylene blue; pH 7.5) [47]. Experiments to the ratiometric UCL imaging of the H 2 O 2 generation were performed in E3 embryo media stimulated with PMA (1 mg/mL) for 20 min at 28.5 C, followed by the microinjection of CYD1-UCNPs (0.5 mg/mL, 20 mL in PBS) into the yolk sac for 120 min.…”

Potentiometric Response of a Neutral-carrier-based Membrane to Aqueous Mercury in Cl−-rich Media

“…1 Various techniques have been reported for mercury detection; these are classified into spectroscopic, radiochemical, and electrochemical methods. 2 The use of ion-selective electrodes (ISEs), one of the most useful electrochemical techniques, has many practical advantages, such as high selectivity, a fast response time, and ease of use.…”

“…More recently, in vivo mercury detection was demonstrated by using Hg 2+ -sensitive fluorescent dyes. 1 We employed a calixarene derivative as an ionophore in conventional liquid polymeric membranes to probe the potentiometric response of neutralcarrier-based ISEs to aqueous mercury species in the presence of a significant amount of Cl -.…”