Two-photon microscopy and its application to live cell imagin

Two-photon fluorescent microscopy is emerging in many areas of science as a premier method of exciting fluorescence in biological samples. This technology enables noninvasive study of biological specimens in three dimensions with improved resolution over conventional imaging methods. Two-photon excitation produces a higher-contrast image and is a novel method to trigger localized photochemical reactions.;In this study, constitutively expressed eGFP in the nucleus of live HeLa cells is monitored over a period of hours. Other conventional imaging methods are analyzed versus a two-photon generated image. An experimental matrix of two-photon settings is used to determine functional settings for two-photon fluorescent microscopy and compare its resolution versus other conventional imaging techniques.