title: Three-dimensional motion tracking for high-resolution optical microscopy, in vivo.
abstract: When conducting optical imaging experiments, in vivo, the signal to noise ratio and effective spatial and temporal resolution is fundamentally limited by physiological motion of the tissue. A three-dimensional (3D) motion tracking scheme, using a multiphoton excitation microscope with a resonant galvanometer, (512 × 512 pixels at 33 frames s(-1)) is described to overcome physiological motion, in vivo. The use of commercially available graphical processing units permitted the rapid 3D cross-correlation of sequential volumes to detect displacements and adjust tissue position to track motions in near real-time. Motion phantom tests maintained micron resolution with displacement velocities of up to 200 μm min(-1), well within the drift observed in many biological tissues under physiologically relevant conditions. In vivo experiments on mouse skeletal muscle using the capillary vasculature with luminal dye as a displacement reference revealed an effective and robust method of tracking tissue motion to enable (1) signal averaging over time without compromising resolution, and (2) tracking of cellular regions during a physiological perturbation.
authors.affiliation: Laboratory of Cardiac Energetics National Heart Lung and Blood Institute, Princeton, New Jersey, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: Bakalar
authors.affiliation: Laboratory of Cardiac Energetics National Heart Lung and Blood Institute, Princeton, New Jersey, USA.
authors.forename: James L
authors.initials: JL
authors.lastname: Schroeder
authors.affiliation: CIT Signal Processing Group, Princeton, New Jersey, USA.
authors.forename: Randall
authors.initials: R
authors.lastname: Pursley
authors.affiliation: CIT Signal Processing Group, Princeton, New Jersey, USA.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Pohida
authors.affiliation: Laboratory of Cardiac Energetics National Heart Lung and Blood Institute, Princeton, New Jersey, USA.
authors.forename: Brian
authors.initials: B
authors.lastname: Glancy
authors.affiliation: Laboratory of Cardiac Energetics National Heart Lung and Blood Institute, Princeton, New Jersey, USA.
authors.forename: Joni
authors.initials: J
authors.lastname: Taylor
authors.affiliation: Laboratory of Cardiac Energetics National Heart Lung and Blood Institute, Princeton, New Jersey, USA.
authors.forename: David
authors.initials: D
authors.lastname: Chess
authors.affiliation: Laboratory of Cardiac Energetics National Heart Lung and Blood Institute, Princeton, New Jersey, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Kellman
authors.affiliation: Siemens Corporate Research, Princeton, New Jersey, USA.
authors.forename: Hui
authors.initials: H
authors.lastname: Xue
authors.affiliation: Laboratory of Cardiac Energetics National Heart Lung and Blood Institute, Princeton, New Jersey, USA.
authors.forename: Robert S
authors.initials: RS
authors.lastname: Balaban
mesh_terms: D000818:Animals; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008124:Locomotion; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods*; D018715:Microscopy, Video / Q000379:methods*; D018482:Muscle, Skeletal / Q000502:physiology*