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Assessment of Cellular Oxidation using a Subcellular Compartment-Specific Redox-Sensitive Green Fluorescent Protein
Published on: June 18, 2020
A colorimetric and fluorescent probe for detecting intracellular biothiols
Chunyang Chen1, Wei Liu1, Cong Xu1
1Key Laboratory of Nonferrous Metals Chemistry and Resources Utilization of Gansu Province and State Key Laboratory of Applied Organic Chemistry, College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou 730000, PR China.
A novel coumarin-based probe enables sensitive and selective detection of intracellular thiols like cysteine and homocysteine. This biocompatible probe facilitates fluorescence imaging in cells and tissues, with potential applications in blood thiol analysis.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Chemical Biology
Background:
- Intracellular thiols play crucial roles in cellular processes.
- Accurate detection of specific thiols like cysteine and homocysteine is vital for understanding biological functions and disease states.
- Existing detection methods often lack sensitivity, selectivity, or biocompatibility for real-time cellular analysis.
Purpose of the Study:
- To design and synthesize a novel, rapid, and highly sensitive coumarin-based fluorescent probe (probe 1) for detecting intracellular thiols.
- To achieve high selectivity for cysteine and homocysteine over other biologically relevant thiols and amino acids under physiological conditions.
- To evaluate the probe's utility for fluorescence imaging and quantitative determination of biothiols in biological samples, including cells, tissues, and serum.
Main Methods:
- Synthesis of probe 1 via a 4-step procedure.
- Evaluation of probe 1's fluorescence response and selectivity towards various thiols and amino acids.
- Application of probe 1 for fluorescence imaging in Hela cells and mice organ tissues.
- Assessment of probe 1's biocompatibility through cell viability assays.
- Determination of thiols in animal serum samples using probe 1.
Main Results:
- Probe 1 demonstrated a latent fluorescence turn-on response with high sensitivity and selectivity for cysteine and homocysteine due to specific cyclization with the aldehyde group.
- Successful fluorescence imaging of intracellular biothiols in Hela cells and quantitative determination within a specific range.
- Demonstrated permeability and specific fluorescence imaging in mice organ tissues.
- Cell viability remained above 80%, indicating good biocompatibility.
- Probe 1 effectively detected thiols in five types of animal serum, showing potential for blood analysis.
Conclusions:
- Probe 1 is a rapid, highly sensitive, and selective coumarin-based fluorescent probe for detecting cysteine and homocysteine.
- The probe exhibits excellent biocompatibility and permeability, enabling effective fluorescence imaging in cells and tissues.
- Probe 1 shows promise as a versatile tool for the quantitative determination of biothiols in various biological matrices, including blood.
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