title: Assessment of automated analyses of cell migration on flat and nanostructured surfaces.
abstract: Motility studies of cells often rely on computer software that analyzes time-lapse recorded movies and establishes cell trajectories fully automatically. This raises the question of reproducibility of results, since different programs could yield significantly different results of such automated analysis. The fact that the segmentation routines of such programs are often challenged by nanostructured surfaces makes the question more pertinent. Here we illustrate how it is possible to track cells on bright field microscopy images with image analysis routines implemented in an open-source cell tracking program, PACT (Program for Automated Cell Tracking). We compare the automated motility analysis of three cell tracking programs, PACT, Autozell, and TLA, using the same movies as input for all three programs. We find that different programs track overlapping, but different subsets of cells due to different segmentation methods. Unfortunately, population averages based on such different cell populations, differ significantly in some cases. Thus, results obtained with one software package are not necessarily reproducible by other software.
authors.affiliation: Department of Micro- and Nanotechnology, Technical University of Denmark, Kgs. Lyngby, Denmark.
authors.forename: Cristian
authors.initials: C
authors.lastname: Grădinaru
authors.affiliation: Department of Micro- and Nanotechnology, Technical University of Denmark, Kgs. Lyngby, Denmark.
authors.forename: Joanna M
authors.initials: JM
authors.lastname: Lopacińska
authors.affiliation: Neural Information Processing, University of Ulm, Ulm, Germany ; Department of Gastroenterology and Endocrinology, University Hospital of Marburg, Marburg, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Huth
authors.affiliation: Neural Information Processing, University of Ulm, Ulm, Germany ; Internal Medicine I - Gastroenterology, University Hospital Ulm, Ulm, Germany.
authors.forename: Hans A
authors.initials: HA
authors.lastname: Kestler
authors.affiliation: Department of Micro- and Nanotechnology, Technical University of Denmark, Kgs. Lyngby, Denmark.
authors.forename: Henrik
authors.initials: H
authors.lastname: Flyvbjerg
authors.affiliation: Department of Micro- and Nanotechnology, Technical University of Denmark, Kgs. Lyngby, Denmark.
authors.forename: Kristian
authors.initials: K
authors.lastname: Mølhave
keywords: Time lapse microscopy; cell migration; cell motility; cell tracking; image analysis; nanotechnology; software