Automated three-dimensional tracking of living cells by digital holographic microscopy.
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.
Langehanenberg
Patrik
P
University Medical Center of Muenster, Center for Biomedical Optics and Photonics, Robert-Koch-Strasse 45, 48149 Muenster, Germany. langehanenberg@uni-muenster.de
Ivanova
Lyubomira
L
Bernhardt
Ingolf
I
Ketelhut
Steffi
S
Vollmer
Angelika
A
Dirksen
Dieter
D
Georgiev
Georgi
G
von Bally
Gert
G
Kemper
Björn
B
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