title: Automated three-dimensional tracking of living cells by digital holographic microscopy.
abstract: Digital holographic microscopy (DHM) enables a quantitative multifocus phase contrast imaging that has been found suitable for technical inspection and quantitative live cell imaging. The combination of DHM with fast and robust autofocus algorithms enables subsequent automated focus realignment by numerical propagation of the digital holographically reconstructed object wave. In combination with a calibrated optical imaging system, the obtained propagation data quantify axial displacements of the investigated sample. The evaluation of quantitative DHM phase contrast images also enables an effective determination of lateral cell displacements. Thus, 3-D displacement data are provided. Results from investigations on sedimenting red blood cells and HT-1080 fibrosarcoma cells in a collagen tissue model demonstrate that DHM enables marker-free automated quantitative dynamic 3-D cell tracking without mechanical focus adjustment.
authors.affiliation: University Medical Center of Muenster, Center for Biomedical Optics and Photonics, Robert-Koch-Strasse 45, 48149 Muenster, Germany. langehanenberg@uni-muenster.de
authors.forename: Patrik
authors.initials: P
authors.lastname: Langehanenberg
authors.forename: Lyubomira
authors.initials: L
authors.lastname: Ivanova
authors.forename: Ingolf
authors.initials: I
authors.lastname: Bernhardt
authors.forename: Steffi
authors.initials: S
authors.lastname: Ketelhut
authors.forename: Angelika
authors.initials: A
authors.lastname: Vollmer
authors.forename: Dieter
authors.initials: D
authors.lastname: Dirksen
authors.forename: Georgi
authors.initials: G
authors.lastname: Georgiev
authors.forename: Gert
authors.initials: G
authors.lastname: von Bally
authors.forename: Björn
authors.initials: B
authors.lastname: Kemper
mesh_terms: D000465:Algorithms; D001185:Artificial Intelligence; D002465:Cell Movement / Q000502:physiology; D002478:Cells, Cultured; D004912:Erythrocytes / Q000166:cytology* / Q000502:physiology*; D006696:Holography / Q000379:methods*; D006801:Humans; D007089:Image Enhancement / Q000379:methods; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008853:Microscopy / Q000379:methods*; D010363:Pattern Recognition, Automated / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012815:Signal Processing, Computer-Assisted