Multiphoton microscopy for monitoring intratissue femtosecond laser surgery effects

Abstract

BACKGROUND AND OBJECTIVE: Multiphoton microscopy/tomography has been used as a novel diagnostic method for corneal imaging with subcellular resolution. Here, we used this technique to monitor femtosecond laser intratissue surgery effects. MATERIALS AND METHODS: Multiphoton microscopy/tomography on rabbits based on intense 90 MHz femtosecond Ti: sapphire laser was realized at intensities of MW-GW/cm(2), whereas the surgical procedures were performed with the same system at a higher light intensity on the order of TW/cm(2). RESULTS: Multiphoton microscopy/tomography proved capable of determining the target of interest, and of visualizing and optically evaluating the in vivo intrastromal laser surgical outcomes with high efficiency. More interesting, using this technique, activated intrastromal keratocytes (myofibroblasts) were detected in vivo 24 hours after intrastromal surgery. CONCLUSION: Multiphoton microscopy/tomography is an efficient and convenient non-invasive imaging method which can be used not only to assess intrastromal microsurgical performance but also to perform in vivo follow-up observations on wound repair.