Coherent scattering in multi-harmonic light microscopy

Biophys J. 2001 Mar;80(3):1568-74. doi: 10.1016/S0006-3495(01)76129-2.


By focusing a pulsed laser beam into a sample, harmonic up-conversion can be generated as well as multi-photon excited fluorescence. Whereas multi-photon excited fluorescence microscopy is well established, the use of multi-harmonic generation for three-dimensional image contrast is very recent. Both techniques can provide similar resolution and, for adequate radiating source density, comparable signal levels, allowing them to be combined in a single versatile instrument. However, harmonic generation differs fundamentally from fluorescence generation in that it is coherent and produces radiation patterns that are highly sensitive to phase. As such, multi-harmonic generation microscopy provides a unique window into molecular spatial organization that is inaccessible to fluorescence.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Membrane / physiology
  • Cell Membrane / ultrastructure*
  • Fluorescent Dyes
  • Hippocampus / cytology
  • Hippocampus / physiology
  • In Vitro Techniques
  • Lasers
  • Light
  • Microscopy, Fluorescence / methods*
  • Pyramidal Cells / cytology*
  • Pyramidal Cells / physiology
  • Pyridinium Compounds
  • Scattering, Radiation
  • Sensitivity and Specificity


  • Fluorescent Dyes
  • Pyridinium Compounds
  • 4-(6-(4-dibutylaminophenyl)-1,3,5-hexatrienyl)-1-(4'-sulfobutyl)pyridinium