Rational design of a FRET-based ratiometric fluorescent probe with large Pseudo-Stokes shift for detecting Hg2+ in living cells based on rhodamine and anthracene fluorophores

Abstract

The development of fluorescent dyes has been a continuing attractive research topic in the field of fluorescence sensing and bioimaging technologies, most of them were subject to a single signal change. In this work, a novel colorimetric and ratiometric fluorescent probe 1 based on rhodamine and anthracene groups was designed and synthesized via the fluorescence resonance energy transfer (FRET) mechanism. Probe 1 showed excellent selectivity, higher sensitivity and ratiometric response to Hg²⁺ in the CH₃CN/H₂O (1/1, v/v) system, with a fast response time (less than 30 s); The fluorescent color changed from purple to orange and the solution visible to the naked-eye changed from colorless to pink. The Pseudo-Stokes shift was 174 nm upon addition of Hg²⁺. The limit of detection (LOD) was calculated to be 0.81 μM and 0.38 μM according to fluorescence and UV/vis measurements, respectively. Furthermore, a possible mechanism for the detection of Hg²⁺ by probe 1 was verified by using ¹H NMR, ESI-MS, and HPLC spectra. Meanwhile, probe 1 was successfully used for cell imaging for the detection of Hg²⁺ in living cells.

Keywords: Cell imaging; FRET; Fluorescent probe; Hg(2+); Ratiometric; Rhodamine.