Characterization of p-bis(O-methylstyryl) benzene as a lifetime and anisotropy decay standard for two-photon induced fluorescence

Joseph R. Lakowicz, Ignacy Gryczynski

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We describe the fluorescence spectral properties of p-bis(O-methylstyryl)benzene (bis-MSB) as a standard for time-resolved measurements of two-photon induced fluorescence. Bis-MSB displays the same single exponential intensity decay in several solvents for one- and two-photon excitation. The anisotropy decay displays the same single correlation time of one- and two-photon excitation. The amplitudes of the anisotropy decay are distinct for one- and two-photon excitation. At some excitation wavelengths the anisotropy amplitude appears to be solely the result of one- and two-photon photoselection, but at shorter wavelengths the anisotropy amplitudes are not related by single constant factor. The absorption range of bis-MSB makes it a suitable standard for one- and two-photon excitation of intrinsic protein fluorescence and extrinsic fluorophores.

Original languageEnglish
Pages (from-to)1-7
Number of pages7
JournalBiophysical Chemistry
Issue number1
Publication statusPublished - 1 Jan 1993



  • Fluorescence intensity decay
  • Protein fluorescence
  • Two-photon induced fluorescence
  • p-Bis(O-methylstyryl)benzene

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