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Monitoring membrane potential with second-harmonic generation
Cold Spring Harbor Protocols
|June 4, 2014
Summary
This study details using second-harmonic generation (SHG) imaging to visualize membrane potential changes in cells. This nonlinear optical method offers a novel approach for electrophysiology research.
Area of Science:
- Nonlinear Optics
- Cellular Electrophysiology
- Biophotonics
Background:
- Second-harmonic generation (SHG) is a nonlinear optical process.
- Monitoring membrane potential is crucial for understanding cellular function.
- Existing fluorescence-based methods have limitations.
Purpose of the Study:
- To describe a protocol for using SHG for imaging membrane potential changes.
- To evaluate SHG as a voltage-sensitive imaging technique.
- To characterize SHG voltage sensitivity in a cellular model.
Main Methods:
- Utilized N1E-115 mouse neuroblastoma cells as a model system.
- Employed styryl and naphthylstyryl dyes (hemicyanines) for membrane staining.
- Performed cell culture, staining, patching, and SHG imaging.
- Characterized SHG intensity's voltage sensitivity and determined optimal laser wavelengths.
Main Results:
- SHG intensities from hemicyanine dyes are sensitive to membrane voltage.
- Precise characterization of SHG voltage sensitivity was achieved.
- Optimal laser fundamental wavelengths for SHG imaging were identified.
Conclusions:
- Second-harmonic generation (SHG) imaging is a viable technique for visualizing membrane potential dynamics.
- This nonlinear optical approach offers unique advantages for cellular electrophysiology.
- The protocol provides a framework for applying SHG imaging to neuronal preparations.
