Multimodal nonlinear optical imaging of Caenorhabditis elegans with multiplex coherent anti-Stokes Raman scattering, third-harmonic generation, second-harmonic generation, and two-photon excitation fluorescence

International audienceMultimodal nonlinear optical microscopy was used to analyze the nematode Caenorhabditis elegans (C. elegans) expressing yellow fluorescent protein (YFP) in the neurons. The lipid and protein distributions were clearly visualized using multiplex coherent anti-Stokes Raman scattering (CARS), second harmonic generation (SHG), third harmonic generation (THG), and two-photon excitation fluorescence (TPEF). The TPEF and SHG signals were found to overlap, which allowed identification of the ventral nerve cord. These findings demonstrate the powerful potential of multimodal nonlinear optical microscopy to reveal C. elegans microstructures in vivo