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Single microbead-based fluorescence "turn on" detection of biothiols by flow cytometry
Ahmed Mohamed1, Xuemeng Li2, Jinquan Li3
1College of Food Science & Technology, Huazhong Agricultural University, Wuhan 430070, China; Department of Biochemistry, Faculty of Agriculture, Benha University, Benha 13736, Egypt.
Talanta
|January 11, 2019
Summary
A novel fluorescent sensor using Polystyrene/Quantum Dots/Gold Nanoparticles (PS/QDs/Au) enables rapid, "turn on" detection of biological thiols (biothiols). This groundbreaking method allows for single microbead analysis and intracellular imaging, advancing biothiol detection capabilities.
Area of Science:
- Biochemistry
- Nanotechnology
- Analytical Chemistry
Background:
- Biological thiols (biothiols), including glutathione (GSH), cysteine (Cys), and homocysteine (Hcy), are crucial for redox homeostasis.
- Dysregulation of biothiols is implicated in various physiological and pathological processes.
- Sensitive and specific detection methods for biothiols are essential for biological and medical research.
Purpose of the Study:
- To develop a novel, highly sensitive fluorescent sensor for the "turn on" detection of biothiols.
- To construct a "Polystyrene/Quantum Dots/Gold Nanoparticles" (PS/QDs/Au) probe for biothiol sensing.
- To demonstrate the capability for single microbead-based detection and intracellular imaging of biothiols.
Main Methods:
- Fabrication of fluorescent microbeads by immobilizing quantum dots (QDs) onto polystyrene (PS) microbeads.
- Quenching of QD fluorescence via gold nanoparticle (Au NP) adsorption through electrostatic interaction.
- Development of a "turn on" fluorescence response mechanism triggered by the presence of biothiols.
- Utilizing flow cytometry for individual microbead signal acquisition and analysis.
Main Results:
- The PS/QDs/Au probe exhibited a "turn on" fluorescence response in the presence of biothiols within 5 minutes.
- Achieved low detection limits: 0.5 μM for GSH, 0.1 μM for Cys, and 0.3 μM for Hcy.
- Successfully demonstrated single microbead-based biothiol detection using flow cytometry for the first time.
- Applied the probe for successful imaging of intracellular biothiols in A549 cells.
Conclusions:
- The developed PS/QDs/Au fluorescent sensor provides a novel and effective platform for rapid "turn on" detection of biothiols.
- The ability to perform single microbead analysis and intracellular imaging highlights the probe's significant potential in biological and biomedical applications.
- This advancement offers a promising tool for studying the roles of biothiols in physiological and disease states.