Title :
Investigating up-conversion fluorescence of Phloxine B
Author_Institution :
Multi-Photon Time-Resolved Imaging Lab., Microcosm Inc., Columbia, MD, USA
Abstract :
The steady-state and time-resolved emission of Phloxine B was examined when excited with 90 fs pulses from a mode-locked Ti:sapphire laser. Above the excitation wavelength (775 nm), Phloxine B was found to display two-photon excitation fluorescence with estimated cross-section for excitation of 0.87×10 -49 cm 4 s/photon, and fluorescence spectrum consistent with one-photon excitation. Over the excitation wavelength range from 590 to 650 nm, Phloxine B was found to display one-photon excitation up-conversion fluorescence. The up-conversion fluorescence of Phloxine B was confirmed by frequency-domain measurements. The intensity decays revealed different double-exponential intensity decays for one-photon excitation at 390 nm and 633 nm. The longer fluorescence lifetimes were observed with 633 nm excitation. Hence, anti-Stokes excitation of Phloxine B causes delay emission at shorter wavelengths. Time-resolved anisotropy decay measurements revealed similar correlation times, but different amplitudes, as has been observed previously for two- versus one-photon excitation.
Keywords :
biochemistry; dyes; fluorescence; molecular biophysics; optical frequency conversion; radiative lifetimes; time resolved spectra; two-photon spectra; 390 nm; 633 nm; Phloxine B; anti-Stokes excitation; double-exponential intensity decays; dye; fluorescence lifetimes; fluorophore; frequency-domain measurements; one-photon excitation up-conversion; steady-state emission; time-resolved anisotropy decay; time-resolved emission; two-photon excitation fluorescence; up-conversion fluorescence; Anisotropic magnetoresistance; Displays; Fluorescence; Frequency measurement; Laser excitation; Laser mode locking; Optical pulses; Propagation delay; Steady-state; Wavelength measurement; Anisotropy; Eosine I Bluish; Ethanol; Fluorescent Dyes; Microscopy, Confocal; Microscopy, Fluorescence; Models, Theoretical;
Journal_Title :
Engineering in Medicine and Biology Magazine, IEEE