Nonlinear microscopy of developing tissues and embryos
Emmanuel Beaurepaire (Lab. for Optics and Biosciences, École Polytechnique/CNRS/INSERM)
Understanding the dynamics at play during embryonic and tissue development requires tissue-scale measurements with subcellular resolution. Nonlinear microscopy is attractive for live studies because it provides tissue 3D imaging and provides several contrast mechanisms. Intrinsic coherent signals such as second- and third-harmonic generation (SHG, THG) provide structural information and can be used in conjunction with fluorescence (2PEF) imaging. SHG signals are specifically obtained from dense organized structures, whereas THG signals are obtained from optical heterogeneities and provide images of the tissue morphology. We will discuss these principles and applications such as imaging the microstructure of intact unstained corneas, and reconstructing the early development of zebrafish embryos. Finally, we will discuss ongoing efforts to improve the capabilities of nonlinear microscopy for imaging developing tissues at advanced stages, such as wavefront control for aberration correction, and simultaneous excitation of multiple fluorescent proteins.
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