Characterization of two-photon point-spread-function in skin imaging applications

摘要

The development of two-photon fluorescence microscopy pushed optical microscopy into a new era. A major advantage of two-photon microscopy is the enhanced depth penetration achievable using near-infrared excitation. This unique feature enables imaging of physiological specimens often to the limit of working distances of high numerical aperture objectives. With the unprecedented capability for in-depth examination of physiological specimens, optical properties of the specimens can affect the imaging formation deep inside the samples. In this work, we address this issue by performing imaging of skin structures by two-photon microscopy. In particular, we like to address the issue of whether objectives with different immersion fluids can influence image parameters. In addition, we will discuss the various methodologies in which the point-spread-function (PSF) may be determined in the skin. Our conclusion is that a direct PSF measurement inside the skin is needed to fully characterize two-photon imaging parameters for optimization of two-photon skin imaging.