Multichannel multiphoton imaging of metal oxides nanoparticles in biological system

Near-IR ultrafast pulse laser and confocal microscope are combined to create a multiphoton multichannel non-linear imaging technique, which allows in situ 3-D characterization of nonfluorescent nanoparticles in biological systems. We observed intense CARS signals generated from various metal oxides due to their high third-order nonlinear susceptibilities (chi((3))), which do not depend on the vibrational resonance but on the electronic resonance. We show that fine and ultrafine particles of metal oxides in alveolar macrophage cells may be imaged in vitro using CARS and multiphoton fluorescence microscopy with highest optical resolution for extended periods without photobleaching effects. The advantage of the epi-detection over the forward detection for imaging sub-micron particles has been investigated.