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Simultaneous Label-Free Autofluorescence Multi-Harmonic Microscopy
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Second Harmonic Imaging of Membrane Potential.

Leslie M Loew1, Aaron Lewis

  • 1Department of Cell Biology, R. D. Berlin Center for Cell Analysis and Modeling, University of Connecticut Health Center, Farmington, CT, 06030-1507, USA, les@volt.uchc.edu.

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|August 5, 2015
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Summary
This summary is machine-generated.

Second Harmonic Imaging Microscopy (SHIM) uses laser scanning to visualize cells. SHIM shows promise for probing cell physiology, but signal strength currently limits imaging fast neuronal electrical activity.

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Area of Science:

  • Non-linear optics
  • Microscopy
  • Cellular imaging

Background:

  • Second Harmonic Generation (SHG) is a non-linear optical effect known since lasers were invented.
  • SHG has emerged as a viable contrast mechanism for microscopy in the last 20 years.
  • SHG microscopy requires minimal modifications to standard laser scanning 2-photon microscopes.

Purpose of the Study:

  • To explore Second Harmonic Imaging Microscopy (SHIM) as a tool for cell and tissue visualization.
  • To investigate the potential of SHIM for probing cell physiology.
  • To assess the limitations of current SHIM technology for imaging dynamic cellular processes.

Main Methods:

  • Utilizing a standard laser scanning 2-photon microscope equipped for SHG imaging.
  • Employing membrane-bound dyes sensitive to membrane potential.
  • Analyzing SHG signal characteristics in relation to cellular activity.

Main Results:

  • SHG signals from specific dyes demonstrate high sensitivity to membrane potential.
  • Current SHIM technology faces limitations due to small signal size.
  • Photon collection during fast action potentials is restricted by signal size.

Conclusions:

  • SHIM is a promising technique for visualizing cell and tissue structure and function.
  • SHIM has the potential to become a valuable probe for cell physiology.
  • Improvements in dyes and microscope optics are needed to image neuronal electrical activity with SHIM.