Third-harmonic microscopy is one of the emerging techniques for noninvasive microscopic imaging of biological structures. We use a novel technique for nonlinear optical material characterization and study the effect of different environment and the structural sensitivity of the third harmonic. In particular, a transformation of collagen in solution is observed for the first time using third-harmonic generation. We also study the ultimate limits of the third harmonic to detect micro- and nanoscopic features inside living cells and find that structures as small as 50 nm can be detected using the current level of technology.
Collagen Imaging, Three-Dimensional