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Highly-Multiplexed Tissue Imaging with Raman Dyes
Published on: April 21, 2022
Nonlinear optical potentiometric dyes optimized for imaging with 1064-nm light
Thomas Z Teisseyre1, Andrew C Millard, Ping Yan
1University of Connecticut Health Center, Richard D. Berlin Center for Cell Analysis and Modeling, 263 Farmington Avenue, MC-1507, Farmington, Connecticut 06030-1507, USA.
Journal of Biomedical Optics
|September 18, 2007
Summary
Researchers developed new voltage-sensitive dyes for deep-tissue imaging of electrical activity. These dyes, optimized for longer wavelengths, show promise for in vivo cellular measurements using nonlinear optical techniques like two-photon fluorescence and second harmonic generation.
Area of Science:
- Biophysics
- Optical Imaging
- Molecular Imaging
Background:
- Nonlinear optical phenomena (two-photon fluorescence and second harmonic generation) combined with voltage-sensitive dyes enable high-resolution mapping of cellular electrical activity.
- Advancements in 1064-nm fiber laser technology allow deep tissue penetration with high-intensity, long-wavelength, femtosecond light pulses.
Purpose of the Study:
- To design and synthesize novel voltage-sensitive dyes optimized for longer wavelengths.
- To screen these new dyes for fast and sensitive responses to membrane potential changes for in vivo applications.
Main Methods:
- Systematic screening of newly synthesized dyes with varied chromophores and membrane-anchoring sidechains.
- Evaluation of dye performance for two-photon fluorescence (2PF) and second harmonic generation (SHG) modalities.
- Analysis of dye responses to membrane potential changes.
Main Results:
- Several novel dyes demonstrated rapid and sensitive responses to membrane potential, suitable for in vivo measurements.
- Some dyes showed optimal performance for SHG, while others were better suited for 2PF.
- Divergent sensitivity and kinetic patterns between SHG and 2PF suggest distinct voltage-sensing mechanisms.
Conclusions:
- The developed dyes hold potential for in vivo imaging of cellular electrical activity.
- The screening approach provides insights into the molecular mechanisms of SHG and 2PF voltage sensitivity.
- Voltage sensitivity in these dyes arises from different mechanisms for SHG and 2PF.
