Browsing by Author "Nelson, Tyler Evan"

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    A Flexible Multimodal 3D Single-Molecule Super-Resolution Microscope for Whole Cell Imaging
    (2023-07-20) Nelson, Tyler Evan; Gustavsson, Anna-Karin
    Single-molecule super resolution techniques can be used to resolve subcellular structures in nanoscale detail, but they are sensitive to background signal which is common in fluorescently labeled cells. Most microscopes are limited to standard epi- illumination, which generates high background fluorescence by illuminating the entire sample at once. Specialized illumination schemes like light sheets or Total Internal Reflection Fluorescence (TIRF) are useful to improve the resolution, but the usefulness of these methods can be limited in certain regions of the cell. In this thesis, we demonstrate a flexible, multimodal super resolution imaging system which combines the optical sectioning capacity of a tilted light sheet with the excellent contrast and homogeneous illumination of a flat-field epi- and TIRF setup. This imaging platform also includes a two-channel 4f point spread function (PSF) engineering system combined with long axial range phase masks for 3D imaging. We show that our microscope greatly reduces background fluorescence throughout thick mammalian cells and improves the performance of single-molecule super-resolution imaging in cells in both 2D and 3D and has the potential to image in 3D throughout an entire cell.