Abstract
Two-photon fluorescence and second harmonics are used to image subsurface structures of highly scattering biological tissues. Ultrashort laser pulses at near-infrared spectra region were reflected by a dichroic beam splitter and then focused into tissue samples by an objective lens to generate nonlinear optical signal. The generated nonlinear optical signal were collected by the same objective lens and passing through the bean splitter to a photo-multiplier tube for detection. Two-photon fluorescence and second harmonics signals were confined in the focal region of the focused beam due to higher order (qudratic) intensity dependence on the excitation light when the focal region is not too deep inside the scattering sample.
© 1999 Optical Society of America
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