Animal model for two-photon excitation in the eye

We wish to deliver Two-Photon Excitation (TPE) to the in vivo retina and to image its effects in order to understand and treat eye disease. A schematic model of the rat eye with a gradient refractive index in the crystalline lens is reconstructed in ZEMAX (TM). This model predicts the monochromatic aberrations as a function of pupil size and field angle and the change in the Point Spread Function (PSF) at best focus. A simple water model of the nonlinear pulse broadening effect has been used to predict the minimal temporal pulse width that will propagate to the retina. In a rat eye uncorrected for monochromatic aberrations, a pupil between 1mm and 1.8mm diameter delivers a peak intensity acceptable for two-photon effects. A somewhat larger diameter pupil (1.35-2.0mm) gives an optimum optical quality for imaging on the optical axis. Correction of the monochromatic aberrations with adaptive optics would improve both imaging and peak intensity. The effect of second order dispersion is dependent on the form of the dispersion relation used. Based on experimental results of second order dispersion, the minimum pulse width to reach the retina is approximately 30fs for the rat eye and approximately 60fs for the human eye.