Journal of Fluorescence

, Volume 3, Issue 2, pp 85–92

Light quenching of tetraphenylbutadiene fluorescence observed during two-photon excitation

  • Ignacy Gryczynski
  • Valery Bogdanov
  • Joseph R. Lakowicz
Article

DOI: 10.1007/BF00865322

Cite this article as:
Gryczynski, I., Bogdanov, V. & Lakowicz, J.R. J Fluoresc (1993) 3: 85. doi:10.1007/BF00865322

Abstract

We observed the steady-state and time-resolved emission of tetraphenylbutadiene (TPB) whea excited by simultaneous absorption of two photons (514 to 610 nm). The intensity initially increased quadratically with laser power, as expected for a two-photon process. At higher laser powers the intensity increases in TPB were subquadratic. The intensity and anisotropy decay times of TPB were unchanged under the locally intense illumination. Importantly, the time zero anisotropy of TPB was decreased under conditions where the intensity was subquadratic. Furthermore, the subquadratic dependence on incident power was not observed for two-photon excitation of 2,5-diphenyloxazole (PPO), for which the incident wavelength does not overlap with the emission spectrum. These results are consistent with stimulated emission (light quenching) of TPB at high laser intensities. The phenomenon of light quenching may be important for other fluorophores used in biochemical research, particularly for the high local intensities used for two-photon excitation.

Key Words

Tetraphenylbutadiene light quenching two-photon excitation 2,5-diphenyloxazole 

Copyright information

© Plenum Publishing Corporation 1993

Authors and Affiliations

  • Ignacy Gryczynski
    • 1
  • Valery Bogdanov
    • 2
  • Joseph R. Lakowicz
    • 1
  1. 1.Center for Fluorescence Spectroscopy and Department of Biological ChemistryUniversity of Maryland School of MedicineBaltimore
  2. 2.Vavilov State Optical InstituteSt. PetersburgRussia

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