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Protein Dynamics in Living Cells

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Simultaneous pH Measurement in Endocytic and Cytosolic Compartments in Living Cells using Confocal Microscopy

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Real-Time Single-Particle Tracking of Intracellular pH Dynamics during Ferroptosis Using Plasmonic Core-Satellite

Hua Liu¹, Xiulin Fan², Lin Wei³

¹School of Pharmaceutical Sciences (Shenzhen), Shenzhen Campus of Sun Yat-Sen University, Shenzhen 518107, China.

|June 12, 2025

Summary

Researchers developed a novel DNA-programmed nanoprobe to map intracellular pH changes during ferroptosis, a cell death process. This tool reveals dynamic pH shifts, aiding the study of cell death mechanisms and cancer therapy development.

Keywords:

ferroptosis mitophagy pH-sensitive plasmon coupling plasmonic core−satellite nanostructures single-particle spectral imaging

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Area of Science:

Biomedical Engineering
Nanotechnology
Cell Biology

Background:

Ferroptosis is an iron-dependent cell death marked by lysosomal changes and pH imbalance.
Understanding intracellular pH dynamics is crucial for studying ferroptosis and developing therapies.

Purpose of the Study:

To develop a novel nanoprobe for real-time, single-particle mapping of intracellular pH during ferroptosis.
To investigate the role of pH dysregulation in ferroptosis and its modulation by mitophagy.

Main Methods:

Fabrication of DNA-programmed gold core-satellite nanostructures (Au CSNSs) with i-motif linkers.
Utilizing pH-dependent conformational changes of i-motif DNA to tune interparticle gaps and LSPR shifts.
Monitoring intracellular pH dynamics in cells undergoing ferroptosis, with and without mitophagy induction.

Main Results:

The Au CSNSs demonstrated reversible and linear pH sensing (R² = 0.97) in the relevant pH range (5.6-6.8).
The nanoprobe successfully mapped rapid intracellular pH changes associated with lysosomal H+ leakage during iron-induced ferroptosis.
Mitophagy activation accelerated and amplified ferroptosis-induced pH alterations.

Conclusions:

A novel DNA-programmed nanoprobe platform enables precise monitoring of intracellular pH dynamics in ferroptosis.
The findings highlight the critical role of pH dysregulation in ferroptosis and provide insights into mitochondria-lysosome crosstalk.
This platform has potential applications in cancer therapy development and understanding cell death pathways.