Research Papers

Automated three-dimensional tracking of living cells by digital holographic microscopy

[+] Author Affiliations
Patrik Langehanenberg

University Medical Center of Muenster, Center for Biomedical Optics and Photonics, Robert-Koch-Strasse 45, 48149 Muenster, Germany

Lyubomira Ivanova, Ingolf Bernhardt

Saarland University, Laboratory of Biophysics, Faculty of Natural and Technical Sciences III, P.O. Box 151150, 66041 Saarbruecken, Germany

Steffi Ketelhut, Angelika Vollmer

University Medical Center of Muenster, Center for Biomedical Optics and Photonics, Robert-Koch-Strasse 45, 48149 Muenster, Germany

Dieter Dirksen

University Medical Center of Muenster, Department of Prosthodontics, Waldeyerstrasse 30, 48149 Muenster, Germany

Georgi Georgiev

St. Klement Ohridski University Sofia, Department of Organic Chemical Technology, Faculty of Chemistry, James Bourchier Boulevard 1, 1164 Sofia, Bulgaria

Gert von Bally, Björn Kemper

University Medical Center of Muenster, Center for Biomedical Optics and Photonics, Robert-Koch-Strasse 45, 48149 Muenster, Germany

J. Biomed. Opt. 14(1), 014018 (February 19, 2009). doi:10.1117/1.3080133
History: Received July 01, 2008; Revised December 09, 2008; Accepted December 17, 2008; Published February 19, 2009
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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.

Figures in this Article
© 2009 Society of Photo-Optical Instrumentation Engineers

Citation

Patrik Langehanenberg ; Lyubomira Ivanova ; Ingolf Bernhardt ; Steffi Ketelhut ; Angelika Vollmer, et al.
"Automated three-dimensional tracking of living cells by digital holographic microscopy", J. Biomed. Opt. 14(1), 014018 (February 19, 2009). ; http://dx.doi.org/10.1117/1.3080133


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