Bioimaging of Glutathione with a Two-Photon Fluorescent Probe and Its Potential Application for Surgery Guide in Laryngeal Cancer

Abstract

As the most abundant nonprotein biothiol in living cells, glutathione (GSH) prevents cellular components from oxidative damage and maintains the intracellular redox homeostasis. For further exploring whether GSH can be employed as a bioindicator to discriminate tumor lesion at a cellular level, the highly selective detection and accurate quantification of GSH under pathological conditions are critical. Herein, we design a coumarin derivative-based two-photon fluorescent probe Cou-Br for the detection of GSH in living cells, mice models, and clinical specimens. The prepared probe is capable of sensitively and selectively detecting GSH in complex biological systems. Cou-Br displays a good linear relationship in response to GSH and a low limit of detection. With the fluorescence signal positively associated with intracellular GSH levels, the probe enables real-time imaging of GSH in various cell lines. Under the condition of CS 2 stimulation, Cou-Br can rapidly respond to the fluctuation of intracellular GSH induced by oxidative damage. Furthermore, the in situ and in vivo bioimaging performances of Cou-Br are demonstrated. Typically, relying on the different cellular concentrations of GSH, the probe is successfully employed to identify the human laryngeal cancer lesion with outstanding capabilities of deep tissue imaging and tumor margin recognition. We assume that the abnormal expression level of GSH may be utilized as a potential bioindicator to discriminate tumor tissues from the surrounding disease-free tissues. To conclude, the proposed probe Cou-Br may potentially serve as a powerful chemical tool for the surgical navigation of cancer in clinic.

Structure modulation on fluorescent probes for biothiols and the reversible imaging of glutathione in living cells

“…The yields were only around 20% to 40%. The probe molecules were fully characterized by 1 H NMR, 13 C NMR and HRMS (Fig. S1-S5 †).…”

“…) 13. C NMR (100 MHz, DMSO-d 6 ) d [ppm]: 187.98, 151.19, 150.82, 150.62, 139.61, 137.86, 135.46, 131.45, 131.27, 131.21, 130.87, 126.67, 125.03, 124.97, 124.59, 124.33, 116.90, 105.29, 40.61.…”

Reduced Thiol Compounds – Induced Biosensing, Bioimaging Analysis and Targeted Delivery

“…With advances in FRET technology, the in vivo study of proteins has gradually developed from surface tissues to deep tissues to the complete detection of amino acids in deep tissue, providing greater accuracy in biological and medicinal research [60] . Tan et al .…”

A red-emission fluorescence probe based on 1,4-addition reaction mechanism for the detection of biothiols in vitro and in vivo

“…Regarding to the detection of biothiols, some methods have been developed, including high-performance liquid chromatography (HPLC), capillary electrophoresis, mass spectrometry (MS) and fluorescence technique. [15][16][17][18][19][20][21] Among them, fluorescent method has been proved to be a desirable tool for in situ biomolecule visualization and detection owing to its simplicity and real-time detection. [22][23][24][25] To date, a large number of fluorescence probes for the detection of biothiols have been reported in the past few years based on various sensing mechanisms: (1) -SH cleavage reactions; (2) -SH nucleophilic reaction; (3) metal-complexes displacement strategies and others.…”