title: Note: Time-gated 3D single quantum dot tracking with simultaneous spinning disk imaging.
abstract: We describe recent upgrades to a 3D tracking microscope to include simultaneous Nipkow spinning disk imaging and time-gated single-particle tracking (SPT). Simultaneous 3D molecular tracking and spinning disk imaging enable the visualization of cellular structures and proteins around a given fluorescently labeled target molecule. The addition of photon time-gating to the SPT hardware improves signal to noise by discriminating against Raman scattering and short-lived fluorescence. In contrast to camera-based SPT, single-photon arrival times are recorded, enabling time-resolved spectroscopy (e.g., measurement of fluorescence lifetimes and photon correlations) to be performed during single molecule/particle tracking experiments.
authors.affiliation: Center for Integrated Nanotechnologies, Los Alamos National Laboratory, Mail Stop G755, Los Alamos, New Mexico 87545, USA.
authors.forename: M S
authors.initials: MS
authors.lastname: DeVore
authors.affiliation: Center for Integrated Nanotechnologies, Los Alamos National Laboratory, Mail Stop G755, Los Alamos, New Mexico 87545, USA.
authors.forename: D G
authors.initials: DG
authors.lastname: Stich
authors.affiliation: Center for Integrated Nanotechnologies, Los Alamos National Laboratory, Mail Stop G755, Los Alamos, New Mexico 87545, USA.
authors.forename: A M
authors.initials: AM
authors.lastname: Keller
authors.affiliation: Department of Pathology and Cancer Research and Treatment Center, University of New Mexico, Albuquerque, New Mexico 87131, USA.
authors.forename: C
authors.initials: C
authors.lastname: Cleyrat
authors.affiliation: Center for Integrated Nanotechnologies, Los Alamos National Laboratory, Mail Stop G755, Los Alamos, New Mexico 87545, USA.
authors.forename: M E
authors.initials: ME
authors.lastname: Phipps
authors.affiliation: Center for Integrated Nanotechnologies, Los Alamos National Laboratory, Mail Stop G755, Los Alamos, New Mexico 87545, USA.
authors.forename: J A
authors.initials: JA
authors.lastname: Hollingsworth
authors.affiliation: Department of Pathology and Cancer Research and Treatment Center, University of New Mexico, Albuquerque, New Mexico 87131, USA.
authors.forename: D S
authors.initials: DS
authors.lastname: Lidke
authors.affiliation: Department of Pathology and Cancer Research and Treatment Center, University of New Mexico, Albuquerque, New Mexico 87131, USA.
authors.forename: B S
authors.initials: BS
authors.lastname: Wilson
authors.affiliation: Center for Integrated Nanotechnologies, Los Alamos National Laboratory, Mail Stop G755, Los Alamos, New Mexico 87545, USA.
authors.forename: P M
authors.initials: PM
authors.lastname: Goodwin
authors.affiliation: Center for Integrated Nanotechnologies, Los Alamos National Laboratory, Mail Stop G755, Los Alamos, New Mexico 87545, USA.
authors.forename: J H
authors.initials: JH
authors.lastname: Werner
mesh_terms: D058948:Cell Tracking / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007089:Image Enhancement / Q000295:instrumentation*; D021621:Imaging, Three-Dimensional / Q000295:instrumentation*; D008856:Microscopy, Fluorescence / Q000295:instrumentation*; D057054:Molecular Imaging / Q000295:instrumentation*; D045663:Quantum Dots; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012815:Signal Processing, Computer-Assisted / Q000295:instrumentation