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Related Experiment Video

Updated: Jun 29, 2025

Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors
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An Ultrasensitive Genetically Encoded Voltage Indicator Uncovers the Electrical Activity of Non-Excitable Cells.

Philipp Rühl1, Anagha G Nair1, Namrata Gawande1

  • 1Center for Molecular Biomedicine, Department of Biophysics, Friedrich Schiller University Jena and Jena University Hospital, D-07745, Jena, Germany.

Advanced Science (Weinheim, Baden-Wurttemberg, Germany)
|March 25, 2024
PubMed
Summary

Scientists developed rEstus, a highly sensitive voltage sensor for non-excitable cells. This tool reveals spontaneous electrical activity and cell-cell communication in previously unstudied cell types.

Keywords:
cellular membrane potentialelectrophysiologyfluorescence imaginggenetically encoded voltage indicatorsnon‐excitable cells

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Area of Science:

  • Cell Biology
  • Neuroscience
  • Biophysics

Background:

  • Non-excitable cells lack action potentials, necessitating highly sensitive voltage sensors.
  • Existing voltage sensors struggle to detect subtle voltage changes in non-excitable cells.

Purpose of the Study:

  • To engineer an ultrasensitive, bright, and calibratable genetically encoded voltage sensor (rEstus).
  • To investigate spontaneous membrane voltage alterations in non-excitable cells.
  • To assess electrical cell-cell coupling using optical voltage recordings.

Main Methods:

  • Structure-guided engineering of a novel genetically encoded voltage sensor, rEstus.
  • Utilizing rEstus for high-sensitivity voltage imaging in non-excitable cell lines (A375, HEK293T, MCF7).
  • Performing correlation analysis on optically recorded voltage signals to determine cell coupling.

Main Results:

  • rEstus exhibits superior brightness and sensitivity in the resting voltage range of non-excitable cells compared to its precursor.
  • Spontaneous endogenous membrane voltage fluctuations were detected in A375, HEK293T, and MCF7 cells on second to millisecond timescales.
  • Direct correlation between visual and electrical coupling was observed in A375 and HEK293T cells, with weaker coupling in MCF7 cells.

Conclusions:

  • rEstus provides an enhanced tool for non-invasive voltage imaging in living non-excitable cells.
  • Spontaneous membrane voltage alterations are a common phenomenon in various non-excitable cell types.
  • Optical voltage recording offers a direct readout of electrical cell-cell communication.