Abstract
By breaking the classical diffraction limit, Stimulated Emission Depletion (STED) Microscopy has revolutionized far-field fluorescence microscopy. 25 nm resolution and better have been achieved in two dimensions imaging cultured cells and even neurons in the brain of living mice. 3D super-resolution has also been demonstrated utilizing two opposing objectives or phase filters with a top-hat profile (see Figure), its application to tissue has however been hampered by aberrations introduced by refractive index inhomogeneities.Here we present our latest results in 3D STED microscopy of scattering specimens enabled by the integration of adaptive optics into a custom STED microscope. We will present our current research about the physical and technical concepts of adaptive optics STED microscopy as well as the latest biological applications of adaptive optics STED microscopy.FigureSchematic of 3D STED Microscopy with aberration correction. A top-hat phase filter (a) is modified to compensate for sample-induced aberrations (b) to achieve an optimized 3D depletion profile (c).View Large Image | View Hi-Res Image | Download PowerPoint Slide
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