title: Simultaneous dual-color 3D STED microscopy.
abstract: We describe the design and implementation of a stimulated emission depletion (STED) microscope which allows simultaneous three-dimensional super-resolution imaging in two colors. A super-continuum laser source is used to provide all spectral bands necessary for excitation and efficient depletion to achieve a lateral and axial resolution of ~35 nm and ~90 nm respectively. We characterize the systems' performance by imaging colloidal particles and single fluorescent molecules. Its biological applicability is demonstrated by dual-color imaging of nuclear pore complexes and of DNA replication sites in mammalian cells.
authors.forename: Christian
authors.initials: C
authors.lastname: Osseforth
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Moffitt
authors.forename: Lothar
authors.initials: L
authors.lastname: Schermelleh
authors.forename: Jens
authors.initials: J
authors.lastname: Michaelis
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D003116:Color; D004247:DNA / Q000378:metabolism; D004261:DNA Replication; D006801:Humans; D021621:Imaging, Three-Dimensional; D051379:Mice; D008853:Microscopy / Q000295:instrumentation / Q000379:methods*; D016196:S Phase
chemical_list: D004247:DNA