Superresolution techniques have pushed the resolution of fluorescence microscopy (FM) towards that of electron microscopy (EM). Meanwhile, developments in scanning EM (SEM) are revolutionizing EM, moving lateral image dimensions to typical FM fields-of-view and extending imaging capability into the third dimension and the live-cell regime. By correlating data from both techniques, molecules can be localized within the context of cells and tissue and with reference to their live dynamics, but throughput and quantification are hindered by elaborate, expert procedures involving separate microscopes. In this presentation, I will show an integrated approach, with high-numerical aperture FM in a SEM, such that the electron beam can be positioned anywhere within the fluorescence field of view[6, 7]. Using electron-beam excited cathodoluminescence from the transparent sample substrate, we achieve automated FM-EM image registration (Fig. 1) with an accuracy that can be pushed to 5 nm, i.e. equalling biomolecular length scales. Besides integrated correlation microscopy, I will show our progress towards novel applications bridging fluorescence and electron microscopy, such as fluorescence-guided live-cell EM, and electron-beam identification and localization of labels, molecules, and cells.
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To cite this abstract:Jacob Hoogenboom; Integrated microscopy: Matching scales and capabilities in light and electron microscopy.. The 16th European Microscopy Congress, Lyon, France. https://emc-proceedings.com/abstract/integrated-microscopy-matching-scales-and-capabilities-in-light-and-electron-microscopy/. Accessed: February 23, 2019
EMC Abstracts - https://emc-proceedings.com/abstract/integrated-microscopy-matching-scales-and-capabilities-in-light-and-electron-microscopy/