Two-photon fluorescence imaging of embryo with much less damage than confocal imaging

Two-photon Laser Scanning Microscopy (TPLSM) is a novel technique based on the two-photon excitation of fluorophore. In this paper, TPLSM and traditional confocal microscopy are introduced. And the influence of femtosecond near-infrared (NIR) illumination on mouse embryos is investigated for the first time. The result shows that NIR laser has much less damage to embryos than blue laser and proves that TPLSM is superior to conventional confocal microscopy in keeping sample alive. TPLSM enables us to make a continuous observation for a longer time on embryogenesis.