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Updated: Jul 29, 2026
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Simultaneous pH Measurement in Endocytic and Cytosolic Compartments in Living Cells using Confocal Microscopy
Published on: April 28, 2014
14.9K
Single-Cell Analysis with Spatiotemporal Control of Local pH
Kelsey Cremin1,2,3,4, Gabriel N Meloni1,2,5, Orkun S Soyer1,4
1Bio-Electrical Engineering Innovation Hub, University of Warwick, Coventry CV4 7AL, United Kingdom.
ACS Measurement Science Au
|February 24, 2025
Summary
This study introduces a new method combining scanning ion conductance microscopy (SICM) and confocal laser scanning microscopy (CLSM) to precisely deliver acid to individual cells. The findings reveal how local pH changes impact cell behavior and recovery, offering new insights for cell biology research.
Area of Science:
- Cell Biology
- Biophysics
- Microscopy Techniques
Background:
- Studying cellular responses to localized environmental changes is crucial.
- Previous methods lacked the precision for single-cell, localized chemical delivery and analysis.
- Understanding pH gradients' impact on cell populations is essential for various biological processes.
Purpose of the Study:
- To develop and validate an integrated platform for precise, localized acid delivery to individual cells within a population.
- To investigate the relationship between extracellular pH gradients and intracellular pH dynamics.
- To analyze the recovery of intracellular pH after localized acid exposure.
Main Methods:
- Integration of scanning ion conductance microscopy (SICM) with confocal laser scanning microscopy (CLSM).
- Utilizing intra- and extracellular pH indicator dyes for real-time monitoring.
- Employing computational modeling to simulate and calculate pH gradients.
- Performing localized acid delivery to individual HeLa cells.
Main Results:
- Demonstrated a strong correlation between imposed extracellular pH gradients and intracellular pH changes.
- Observed that severe acid challenges prevent full intracellular pH recovery even after extracellular pH normalization.
- Highlighted the localized nature of the proton gradient due to media buffering.
Conclusions:
- The combined SICM-CLSM platform enables precise, localized chemical delivery and single-cell analysis within populations.
- Intracellular pH is highly sensitive to localized extracellular pH gradients.
- This methodology provides a powerful tool for studying cellular responses to controlled microenvironmental conditions.
