3D tracking and multi-wavelength techniques for digital holographic microscopy based cell analysis

Björn Kemper; Patrik Langehanenberg; Sebastian Kosmeier; Sabine Przibilla; Angelika Vollmer; Steffi Ketelhut; Gert von Bally

doi:10.1117/12.831552

1 July 2009 3D tracking and multi-wavelength techniques for digital holographic microscopy based cell analysis

Björn Kemper, Patrik Langehanenberg, Sebastian Kosmeier, Sabine Przibilla, Angelika Vollmer, Steffi Ketelhut, Gert von Bally

Author Affiliations +

Proceedings Volume 7367, Advanced Microscopy Techniques; 736714 (2009) https://doi.org/10.1117/12.831552
Event: European Conferences on Biomedical Optics, 2009, Munich, Germany

Abstract

Digital Holographic Microscopy (DHM) allows quantitative multi-focus 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 and a calibrated imaging system enables the determination of axial sample displacements. The evaluation of quantitative DHM phase contrast images permits also an effective detection of lateral object movements. Thus, data for 3D tracking is provided. Multi-wavelength techniques open up prospects for an increased phase resolution in DHM by reduction of parasitic interference effects due to multiple reflections within the measurement setup. For this purpose, the generation of short coherence properties by tunable laser light has been investigated for application in DHM. Results from investigations on sedimenting erythrocytes in suspension demonstrate that DHM enables (automated) quantitative dynamic 3D tracking of multiple cells without mechanical focus adjustment. Furthermore, it is shown that multi-wavelength techniques enhance the phase resolution in quantitative digital holographic cell imaging.

Citation Download Citation

Björn Kemper, Patrik Langehanenberg, Sebastian Kosmeier, Sabine Przibilla, Angelika Vollmer, Steffi Ketelhut, and Gert von Bally "3D tracking and multi-wavelength techniques for digital holographic microscopy based cell analysis", Proc. SPIE 7367, Advanced Microscopy Techniques, 736714 (1 July 2009); https://doi.org/10.1117/12.831552

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KEYWORDS

Digital holography

Holograms

Holography

Microscopy

Phase contrast

Microscopes

3D image reconstruction

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