Effect of pulse duration on two-photon excited fluorescence and second harmonic generation in nonlinear optical microscopy.

Publication Type:

Journal Article

Source:

Journal of biomedical optics, Volume 11, Issue 2, p.020501 (2006)

Keywords:

Algorithms, Equipment Design, Equipment Failure Analysis, Humans, Image Enhancement, Microscopy, Fluorescence, Multiphoton, Nonlinear Dynamics, Reproducibility of Results, Sensitivity and Specificity, Signal Processing, Computer-Assisted, Skin

Abstract:

We have developed a multiphoton microscopy (MPM) system using a 12-fs Ti:sapphire laser with adjustable dispersion precompensation in order to examine the impact of pulse duration on nonlinear optical signals. The efficiencies of two-photon-excited fluorescence (TPEF) and second harmonic generation (SHG) were studied for various pulse durations, measured at the sample, ranging from approximately 400 fs to sub-20 fs. Both TPEF and SHG increased proportionally to the inverse of the pulse duration for the entire tested range. Because of improved signal-to-noise ratio, sub-20-fs pulses were used to enhance MPM imaging depth by approximately 160%, compared to 120-fs pulses, in human skin.

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