Single-Cell Analysis of Zinc Ions During Electrical Stimulation-Induced Pyroptosis by DNAzyme-Functionalized Glass...

Area of Science:

• Biomedical Engineering
• Analytical Chemistry
• Cell Biology

Background:

• Zinc ions (Zn²⁺) are essential redox elements involved in critical cellular functions like immune response and signal regulation.
• Understanding intracellular Zn²⁺ dynamics is vital for elucidating physiological processes and disease mechanisms.
• Existing methods for intracellular ion detection often face challenges with precision and cell membrane integrity.

Purpose of the Study:

• To develop and validate a novel sensing platform for real-time, intracellular detection of Zn²⁺.
• To utilize DNAzyme-functionalized glass nanopipettes (D-GNs) for sensitive and specific Zn²⁺ measurement within single cells.
• To explore the application of D-GNs in monitoring cellular responses to stimuli.

Main Methods:

• Fabrication of glass nanopipettes functionalized with DNAzymes.
• Utilizing the catalytic activity of Zn²⁺-dependent DNAzymes for signal generation.
• Measuring changes in ionic current rectification due to Zn²⁺-mediated release of DNAzyme-modified gold nanoparticles (D-AuNPs).
• In vitro validation and application in live HeLa cells under electrical stimulation.

Main Results:

• D-GNs demonstrated effective sensing performance in vitro.
• The platform successfully detected intracellular Zn²⁺ in HeLa cells.
• Electrical stimulation induced measurable changes in intracellular Zn²⁺ levels detected by D-GNs.
• The method showed minimal damage to the cell membrane, facilitating single-cell analysis.

Conclusions:

• DNAzyme-functionalized glass nanopipettes provide a sensitive and minimally invasive platform for intracellular Zn²⁺ detection.
• This technology offers a new strategy for probing dynamic changes in intracellular ion concentrations.
• The D-GNs platform holds promise for evaluating the physiological state of individual cells and understanding ion-related cellular processes.