Hyper-Rayleigh and hyper-Raman scattering background of liquid water in two-photon excited fluorescence detection

Anal Chem. 1997 Apr 1;69(7):1285-7. doi: 10.1021/ac961185o.

Abstract

The detection of two-photon excited fluorescence is practically free from background caused by linear scattering because two-photon excited fluorescence occurs at a much shorter wavelength region than the excitation light. This property can be used to achieve ultrasensitive fluorescence detection. However, similar to linear scattering in the detection of one-photon excited fluorescence, the question arises whether background caused by non-linear scattering may limit the detection sensitivity of two-photon excited fluorescence. In this work, quantitative comparisons between two-photon induced scattering of liquid water and two-photon excited dye fluorescence in a standard epifluorescence geometry show that the relative scattering background is typically reduced by orders of magnitude in two-photon excitation as compared to single-photon excitation with confocal detection. Hyper-Rayleigh and hyper-Raman (3400 cm-1) cross sections of liquid water have been measured to be 8 x 10(-62) and 7 x 10(-63) cm4.s/photon, respectively, at 840 nm incident wavelength, with absolute values calibrated with respect to the known two-photon fluorescence excitation cross section of fluorescein.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Fluorescence
  • Lasers
  • Photons
  • Scattering, Radiation
  • Spectrum Analysis, Raman
  • Water / chemistry*

Substances

  • Water