title: Deconvolution-free Subcellular Imaging with Axially Swept Light Sheet Microscopy.
abstract: The use of propagation invariant Bessel beams has enabled high-resolution subcellular light sheet fluorescence microscopy. However, the energy within the concentric side lobe structure of Bessel beams increases significantly with propagation length, generating unwanted out-of-focus fluorescence that enforces practical limits on the imaging field of view size. Here, we present a light sheet fluorescence microscope that achieves 390 nm isotropic resolution and high optical sectioning strength (i.e., out-of-focus blur is strongly suppressed) over large field of views, without the need for structured illumination or deconvolution-based postprocessing. We demonstrate simultaneous dual-color, high-contrast, and high-dynamic-range time-lapse imaging of migrating cells in complex three-dimensional microenvironments, three-dimensional tracking of clathrin-coated pits, and long-term imaging spanning >10 h and encompassing >2600 time points.
authors.affiliation: Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, Texas.
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Dean
authors.affiliation: Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, Texas.
authors.forename: Philippe
authors.initials: P
authors.lastname: Roudot
authors.affiliation: Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, Texas.
authors.forename: Erik S
authors.initials: ES
authors.lastname: Welf
authors.affiliation: Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, Texas.
authors.forename: Gaudenz
authors.initials: G
authors.lastname: Danuser
authors.affiliation: Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, Texas. Electronic address: reto.fiolka@utsouthwestern.edu.
authors.forename: Reto
authors.initials: R
authors.lastname: Fiolka
mesh_terms: D018929:Cell Culture Techniques; D002465:Cell Movement; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000295:instrumentation / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D061848:Optical Imaging / Q000295:instrumentation / Q000379:methods*; D055213:Retinal Pigment Epithelium / Q000648:ultrastructure*; D059008:Time-Lapse Imaging / Q000295:instrumentation / Q000379:methods*
chemical_list: D049452:Green Fluorescent Proteins