Nile-Red-Based Fluorescence Probe for Selective Detection of Biothiols, Computational Study, and Application in Cell
Xiang Rong1, Zhong-Yong Xu1, Jin-Wu Yan1
1MOE International Joint Research Laboratory on Synthetic Biology and Medicines, School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, China.
Molecules (Basel, Switzerland)
|October 17, 2020
Summary
A novel Nile-red based probe, NRSH, enables sensitive colorimetric and fluorescent detection of biothiols. This probe offers rapid response and low detection limits, proving effective for real-time bioimaging in cells.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Chemical Biology
Background:
- Biothiols play crucial roles in various biological processes.
- Accurate detection of biothiols is essential for understanding cellular functions and disease mechanisms.
- Development of selective and sensitive probes for biothiols is an ongoing challenge.
Purpose of the Study:
- To develop a new colorimetric and fluorescence probe (NRSH) for selective biothiol detection.
- To investigate the probe's response characteristics, including selectivity, response time, and detection limits.
- To evaluate the probe's applicability for bioimaging of biothiols in living cells.
Main Methods:
- Synthesis of the NRSH probe based on the Nile-red chromophore.
- Spectroscopic analysis (UV-Vis absorption and fluorescence) to characterize probe-biothiol interactions.
- Evaluation of probe selectivity against common interfering species.
- Determination of detection limits for various biothiols (H2S, Cys, GSH, Hcy).
- Cytotoxicity assays and live-cell bioimaging experiments using HeLa cells.
Main Results:
- The NRSH probe exhibited a significant blue shift in absorption (587 nm to 567 nm) and a 35-fold fluorescence enhancement (at 645 nm) upon reaction with biothiols.
- The probe demonstrated high selectivity for biothiols over other species.
- NRSH showed a rapid response (<1 min) for H2S and low detection limits (nM range) for H2S, Cys, GSH, and Hcy.
- The probe displayed low cytotoxicity and was successfully used for real-time biothiol monitoring in HeLa cells via fluorescence imaging.
Conclusions:
- The developed NRSH probe is a highly selective and sensitive tool for colorimetric and fluorescent detection of biothiols.
- Its rapid response, low detection limits, and suitability for live-cell imaging make it valuable for biological and biomedical research.
- Theoretical calculations further elucidated the fluorescence mechanism of the NRSH probe.
