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Simultaneous intracellular chloride and pH measurements using a GFP-based sensor.

Daniele Arosio1, Fernanda Ricci, Laura Marchetti

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Summary

Researchers created ClopHensor, a biosensor for tracking pH and chloride levels in cells. This tool detected high chloride concentrations within cellular granules, advancing live-cell imaging capabilities.

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Area of Science:

  • Cellular Biology
  • Biochemistry
  • Genetics

Background:

  • Chloride ions and protons (pH) play crucial, interconnected roles in cellular signaling and responses.
  • Existing methods for simultaneously monitoring these analytes in real-time within live cells are limited.

Purpose of the Study:

  • To develop a novel ratiometric biosensor for the simultaneous, real-time optical detection of pH and chloride fluxes.
  • To characterize the biosensor's performance in live cellular environments.

Main Methods:

  • Development of a ratiometric biosensor, ClopHensor, utilizing a chloride-sensitive Aequorea victoria Green Fluorescent Protein (GFP) variant.
  • Targeting ClopHensor to specific intracellular compartments, such as large dense-core exocytosis granules.

Main Results:

  • The ClopHensor biosensor successfully enabled combined real-time optical detection of pH changes and chloride fluxes.
  • High chloride concentrations were detected within large dense-core exocytosis granules using the targeted biosensor.

Conclusions:

  • ClopHensor is a valuable tool for simultaneous real-time monitoring of intracellular pH and chloride dynamics.
  • The biosensor facilitates the study of chloride's role in cellular processes, including exocytosis.