title: Three-dimensional reconstruction and motion analysis of living, crawling cells.
abstract: Cell behavior is three-dimensional (3-D), even when it takes place on a flat surface. Migrating cells form pseudopods on and off the substratum, and the cell body undergoes height changes associated with a 1 min behavior cycle. Inside the cell, the nucleus has a 3-D migratory cycle, and vesicles move up and down in the z-axis as a cell locomotes. For these reasons, the two-dimensional (2-D) analysis of cellular and subcellular behavior is, in many cases, inadequate. We have, therefore, developed 3-D motion analysis systems that reconstruct the cell surface, nucleus, pseudopods, and vesicles of living, crawling cells in 3-D at time intervals as short as 1 s, and compute more than 100 parameters of motility and dynamics morphology at 1-s intervals. We are now in the process of developing a multimode reconstruction system that will allow us to reconstruct and analyze fluorescently tagged molecular complexes within the differential interference contrast-imaged subcellular architecture of a crawling cell. These evolving technologies should find wide application for a host of biomedical problems.
authors.affiliation: W.M. Keck Dynamic Image Analysis Facility, Department of Biological Sciences, The University of Iowa, Iowa City 52242, USA. david-soll@uiowa.edu
authors.forename: D R
authors.initials: DR
authors.lastname: Soll
authors.forename: E
authors.initials: E
authors.lastname: Voss
authors.forename: O
authors.initials: O
authors.lastname: Johnson
authors.forename: D
authors.initials: D
authors.lastname: Wessels
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000502:physiology / Q000648:ultrastructure; D002465:Cell Movement; D002467:Cell Nucleus / Q000502:physiology / Q000648:ultrastructure; D004023:Dictyostelium / Q000502:physiology* / Q000648:ultrastructure*; D007091:Image Processing, Computer-Assisted; D008857:Microscopy, Interference / Q000295:instrumentation* / Q000379:methods; D008858:Microscopy, Phase-Contrast / Q000295:instrumentation* / Q000379:methods; D015388:Organelles / Q000502:physiology / Q000648:ultrastructure; D011554:Pseudopodia / Q000502:physiology / Q000648:ultrastructure; D012984:Software; D014584:User-Computer Interface