Fluorescent Probes for Live Cell Thiol Detection

Thiols play vital and irreplaceable roles in the biological system. Abnormality of thiol levels has been linked with various diseases and biological disorders. Thiols are known to distribute unevenly and change dynamically in the biological system. Methods that can determine thiols’ concentration and distribution in live cells are in high demand. In the last two decades, fluorescent probes have emerged as a powerful tool for achieving that goal for the simplicity, high sensitivity, and capability of visualizing the analytes in live cells in a non-invasive way. They also enable the determination of intracellular distribution and dynamitic movement of thiols in the intact native environments. This review focuses on some of the major strategies/mechanisms being used for detecting GSH, Cys/Hcy, and other thiols in live cells via fluorescent probes, and how they are applied at the cellular and subcellular levels. The sensing mechanisms (for GSH and Cys/Hcy) and bio-applications of the probes are illustrated followed by a summary of probes for selectively detecting cellular and subcellular thiols.

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Novel anthracene- and pyridine-containing Schiff base probe for selective “off–on” fluorescent determination of Cu2+ ions towards live cell application

New Journal of Chemistry ◽

10.1039/c5nj03168k ◽

2016 ◽

Vol 40 (7) ◽

pp. 6101-6108 ◽

Cited By ~ 14

Author(s):

Turibius Simon ◽

Muthaiah Shellaiah ◽

Venkatesan Srinivasadesikan ◽

Ching-Chang Lin ◽

Fu-Hsiang Ko ◽

...

Keyword(s):

Electron Transfer ◽

Schiff Base ◽

Photoinduced Electron Transfer ◽

Transfer Mechanism ◽

Live Cell ◽

Live Cells ◽

Electron Transfer Mechanism

A simple anthracene-based AP probe was synthesized to detect Cu2+ ions, via the photoinduced electron transfer mechanism, in live cells.

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Simple and efficient delivery of cell-impermeable organic fluorescent probes into live cells for live-cell superresolution imaging

Light Science & Applications ◽

10.1038/s41377-019-0188-0 ◽

2019 ◽

Vol 8 (1) ◽

Cited By ~ 9

Author(s):

Meng Zhang ◽

Meihua Li ◽

Wenting Zhang ◽

Yubing Han ◽

Yu-Hui Zhang

Keyword(s):

Fluorescent Probes ◽

Live Cell ◽

Live Cells ◽

Superresolution Imaging ◽

Efficient Delivery

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Dual-color Fluorescence Cross-Correlation Spectroscopy to study Protein-Protein Interaction and Protein Dynamics in Live Cells

10.1101/2021.09.10.459760 ◽

2021 ◽

Author(s):

Katherina Hemmen ◽

Susobhan Choudhury ◽

Mike Friedrich ◽

Johannes Balkenhol ◽

Felix Knote ◽

...

Keyword(s):

Conformational Changes ◽

High Reliability ◽

Resonance Energy ◽

High Sensitivity ◽

Correlation Spectroscopy ◽

Live Cell ◽

Fluorescence Labeling ◽

Live Cells ◽

Noise Levels ◽

Dual Color

We present a protocol and workflow to perform live cell dual-color fluorescence crosscorrelation spectroscopy (FCCS) combined with Förster Resonance Energy transfer (FRET) to study membrane receptor dynamics in live cells using modern fluorescence labeling techniques. In dual-color FCCS, where the fluctuations in fluorescence intensity represents the dynamical "fingerprint" of the respective fluorescent biomolecule, we can probe co-diffusion or binding of the receptors. FRET, with its high sensitivity to molecular distances, serves as a well-known "nanoruler" to monitor intramolecular changes. Taken together, conformational changes and key parameters such as local receptor concentrations, and mobility constants become accessible in cellular settings. Quantitative fluorescence approaches are challenging in cells due to high noise levels and the vulnerable sample itself. We will show how to perform the experiments including the calibration steps. We use dual-color labeled β2-adrenergic receptor (β2AR) labeled (eGFP and SNAPtag-TAMRA). We will guide you step-by-step through the data analysis procedure using open-source software and provide templates that are easy to customize. Our guideline enables researchers to unravel molecular interactions of biomolecules in live cells in situ with high reliability despite the limited signal-to-noise levels in live cell experiments. The operational window of FRET and particularly FCCS at low concentrations allows quantitative analysis near-physiological conditions.

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Michael addition-based probes for ratiometric fluorescence imaging of protein S-depalmitoylases in live cells and tissues

Chemical Science ◽

10.1039/c7sc02805a ◽

2017 ◽

Vol 8 (11) ◽

pp. 7588-7592 ◽

Cited By ~ 17

Author(s):

Michael W. Beck ◽

Rahul S. Kathayat ◽

Candace M. Cham ◽

Eugene B. Chang ◽

Bryan C. Dickinson

Keyword(s):

Fluorescence Imaging ◽

Fluorescent Probes ◽

Michael Addition ◽

Enzyme Activities ◽

Protein S ◽

Live Cell ◽

Tissue Imaging ◽

Live Cells ◽

Ratiometric Fluorescence

Ratiometric fluorescent probes for cysteine palmitoylation “erasers” permit live cell and tissue imaging of endogenous enzyme activities.

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Lysosome-targeted two-photon fluorescent probes for rapid detection of H2S in live cells

New Journal of Chemistry ◽

10.1039/c8nj05419c ◽

2019 ◽

Vol 43 (3) ◽

pp. 1267-1274 ◽

Cited By ~ 17

Author(s):

Gongchun Li ◽

Saige Ma ◽

Jun Tang ◽

Yong Ye

Keyword(s):

Fluorescent Probes ◽

Rapid Detection ◽

High Sensitivity ◽

Live Cells ◽

Two Photon

Two novel two-photon fluorescent probes (BHNP-DA and M2) were designed and synthesized and show good selectivity and high sensitivity to H2S.

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Light up ClO−in live cells using an aza-coumarin based fluorescent probe with fast response and high sensitivity

The Analyst ◽

10.1039/c5an00777a ◽

2015 ◽

Vol 140 (13) ◽

pp. 4594-4598 ◽

Cited By ~ 54

Author(s):

Jiangli Fan ◽

Huiying Mu ◽

Hao Zhu ◽

Jingyun Wang ◽

Xiaojun Peng

Keyword(s):

Fluorescent Probe ◽

Live Cell Imaging ◽

Cell Imaging ◽

High Sensitivity ◽

Fast Response ◽

Live Cell ◽

Live Cells

An aza-coumarin based fluorescent and colorimetricAC-ClOfor the ClO−determination with fast response and high sensitivity.AC-ClOwas successfully applied for the live-cell imaging of exogenous and endogenous ClO−.

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New Approaches to the Diagnosis of Liver Fibrosis in Patients with Chronic Hepatitis C

Science and Innovations in Medicine ◽

10.35693/2500-1388-2018-0-2-15-18 ◽

2018 ◽

Vol 3 (2) ◽

pp. 15-18

Author(s):

SYu Yu Vasilev ◽

DYu Yu Konstantinov ◽

LL L Popova

Keyword(s):

Chronic Hepatitis ◽

Hepatitis C ◽

Liver Fibrosis ◽

Chronic Hepatitis C ◽

High Sensitivity ◽

Linear Discriminant ◽

Discriminant Model ◽

Non Invasive ◽

Chronic Hcv

Aim - to develop the method of non-invasive determination of liver fibrosis in patients with chronic hepatitis С. Materials and methods. 446 patients with chronic HCV infection (aged from 18 to 65 years; male -52.3%; female - 47.7%) were comprehensively surveyed. The linear discriminant analysis with the exception of discriminatory variables used by the all registered laboratory and ultrasound indicators, as well as the age of patients was performed. The results of the survey of 223 patients were used to create the discriminant model and the results of a survey of the remaining 223 patients, were used for the testing of the accuracy of the received discrimination. Conclusion. The discriminant model for noninvasive determination of the degree of liver fibrosis in patients with chronic hepatitis C with high sensitivity - 88.0% and specificity is 96.2% is developed.

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