Microfluorometric detection of catecholamines with multiphoton-excited fluorescence

Appl Opt. 2004 Apr 20;43(12):2412-7. doi: 10.1364/ao.43.002412.

Abstract

We demonstrate sensitive spatially resolved detection of physiological chromophores that emit in the ultraviolet (<330 nm). An atypical laser source (a visible wavelength femtosecond optical parametric oscillator), and an unconventional collection geometry (a lensless detector that detects the forward-emitted fluorescence) enable this detection. We report the excitation spectra of the catecholamines dopamine and norepinephrine, together with near-UV emitters serotonin and tryptophan, in the range of 550-595 nm. We estimate the molecular two-photon action cross section of dopamine, norepinephrine, and serotonin to be 1.2 mGM (1 GM, or Goppert Mayor, is equal to 10(-58) m4 s(-1) photon(-1)), 2 mGM, and 43 mGM, respectively, at 560 nm. The sensitivity achieved by this method holds promise for the microscopic imaging of vesicular catecholamines in live cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Catecholamines / analysis*
  • Dopamine / analysis
  • Epinephrine / analysis
  • Fluorescence*
  • Fluorometry*
  • Photons*
  • Sensitivity and Specificity
  • Serotonin / analysis
  • Tryptophan / analysis

Substances

  • Catecholamines
  • Serotonin
  • Tryptophan
  • Dopamine
  • Epinephrine