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Label-Free Imaging of Membrane Potentials by Intramembrane Field Modulation, Assessed by Second Harmonic Generation

Yovan de Coene1, Stijn Jooken2, Olivier Deschaume2

  • 1Laboratory of Molecular Imaging and Photonics, Department of Chemistry, KU Leuven, Celestijnenlaan 200D, Leuven, 3001, Belgium.

Small (Weinheim an Der Bergstrasse, Germany)
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Summary

Label-free optical imaging using endogenous second-harmonic generation (SHG) can detect cellular electrical activity. This technique shows high sensitivity to changes in transmembrane potential (TMP) in live cells without external probes.

Keywords:
label-free imagingmembrane potentialsnonlinear opticssecond-harmonic generation

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Area of Science:

  • Biophysics
  • Cellular Electrophysiology
  • Nonlinear Optics

Background:

  • Optical methods are crucial for studying cellular electrical activity.
  • Voltage-sensitive dyes, while useful, can alter cell function due to their lipophilic nature.
  • Label-free techniques avoid exogenous probes, offering a significant advantage.

Purpose of the Study:

  • To investigate the sensitivity of endogenous second-harmonic generation (SHG) to transmembrane potential (TMP) in live cells.
  • To establish a label-free method for monitoring cellular electrical activity.
  • To provide a theoretical framework for SHG voltage sensitivity.

Main Methods:

  • Utilized whole-cell voltage-clamp electrophysiology on human embryonic kidney (HEK293T) cells.
  • Measured endogenous second-harmonic generation (SHG) intensity from live cells.
  • Correlated SHG intensity with controlled changes in transmembrane potential (TMP).

Main Results:

  • Demonstrated a linear relationship between SHG intensity and membrane voltage in HEK293T cells.
  • Observed high optical sensitivity to membrane voltage (approximately 40% per 100 mV).
  • Ruled out membrane hydration as the primary factor for SHG voltage sensitivity.

Conclusions:

  • Endogenous SHG is a sensitive, label-free optical probe for cellular transmembrane potential.
  • The observed sensitivity arises from the electric field within the asymmetric lipid bilayer.
  • SHG offers a promising non-invasive tool for monitoring electrogenic cells.