title: Fast Three-Dimensional Single-Particle Tracking in Natural Brain Tissue.
abstract: Observation of molecular dynamics is often biased by the optical very heterogeneous environment of cells and complex tissue. Here, we have designed an algorithm that facilitates molecular dynamic analyses within brain slices. We adjust fast astigmatism-based three-dimensional single-particle tracking techniques to depth-dependent optical aberrations induced by the refractive index mismatch so that they are applicable to complex samples. In contrast to existing techniques, our online calibration method determines the aberration directly from the acquired two-dimensional image stream by exploiting the inherent particle movement and the redundancy introduced by the astigmatism. The method improves the positioning by reducing the systematic errors introduced by the aberrations, and allows correct derivation of the cellular morphology and molecular diffusion parameters in three dimensions independently of the imaging depth. No additional experimental effort for the user is required. Our method will be useful for many imaging configurations, which allow imaging in deep cellular structures.
authors.affiliation: Research Group for Molecular Physiology, Leibniz Institute for Neurobiology, Magdeburg, Germany; Research Group for Image Processing and Pattern Recognition, Otto-von-Guericke University, Magdeburg, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Sokoll
authors.affiliation: Special Lab for Electron and Laserscanning Microscopy, Leibniz Institute for Neurobiology, Magdeburg, Germany.
authors.forename: Yury
authors.initials: Y
authors.lastname: Prokazov
authors.affiliation: Research Group for Molecular Physiology, Leibniz Institute for Neurobiology, Magdeburg, Germany.
authors.forename: Magnus
authors.initials: M
authors.lastname: Hanses
authors.affiliation: Research Group for Molecular Physiology, Leibniz Institute for Neurobiology, Magdeburg, Germany; Institute of Neural and Sensory Physiology, Medical Faculty, University of Düsseldorf, Germany.
authors.forename: Barbara
authors.initials: B
authors.lastname: Biermann
authors.affiliation: Research Group for Image Processing and Pattern Recognition, Otto-von-Guericke University, Magdeburg, Germany.
authors.forename: Klaus
authors.initials: K
authors.lastname: Tönnies
authors.affiliation: Research Group for Molecular Physiology, Leibniz Institute for Neurobiology, Magdeburg, Germany. Electronic address: martin.heine@lin-magdeburg.de.
authors.forename: Martin
authors.initials: M
authors.lastname: Heine
mesh_terms: D000465:Algorithms; D000818:Animals; D001921:Brain / Q000166:cytology / Q000378:metabolism*; D002138:Calibration; D004058:Diffusion; D021621:Imaging, Three-Dimensional / Q000379:methods*; D051379:Mice; D008959:Models, Neurological; D056004:Molecular Dynamics Simulation; D057054:Molecular Imaging / Q000379:methods*; D013995:Time; D046509:Tissue Culture Techniques / Q000379:methods*