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Second-harmonic generation sensitivity to transmembrane potential in normal and tumor cells.
L Sacconi1, M D'Amico, F Vanzi
1University of Trento, Department of Physics, via Sommarive 14, I-38050 Povo, Trento, Italy.
Journal of Biomedical Optics
|May 25, 2005
Summary
Second-harmonic generation (SHG) offers sensitive optical measurement of cell transmembrane potential. This technique shows higher sensitivity in tumor cells and does not alter cell electrophysiology, making it valuable for neuroscience research.
Area of Science:
- Biophysics
- Cell Biology
- Optical Physics
Background:
- Second-harmonic generation (SHG) is an emerging optical technique.
- It offers high sensitivity and temporal resolution for measuring cellular properties.
- Transmembrane potential is a critical indicator of cell function.
Purpose of the Study:
- To characterize the sensitivity of SHG signals to transmembrane potential.
- To compare SHG sensitivity across different normal and tumor cell types.
- To validate SHG as a non-invasive tool for electrophysiology.
Main Methods:
- Utilized patch clamp techniques to measure transmembrane potential.
- Employed the RH 237 dye for fluorescent labeling of cell membranes.
- Quantified SHG signal changes in response to transmembrane potential variations.
Main Results:
- SHG sensitivity to transmembrane potential varied significantly by cell type (10-17% per 100 mV).
- Tumor cell lines exhibited higher SHG sensitivity than normal cells.
- SV40-induced transformation increased SHG sensitivity in Balb/c3T3 cells.
- RH 237 labeling did not affect cell electrophysiological properties.
Conclusions:
- SHG is a sensitive and non-invasive method for optical measurement of transmembrane potential.
- The technique's sensitivity is modulated by cell type and transformation status.
- SHG holds promise for advancing electrophysiology and neurobiology research.