Special Section on Selected Topics in Biophotonics: Photodynamic Therapy and Optical Micromanipulation for Biophotonics

Contrast-enhanced digital holographic imaging of cellular structures by manipulating the intracellular refractive index

[+] Author Affiliations
Christina E. Rommel, Christian Dierker

University of Muenster, Department of Medicine B, Gastroenterological Molecular Cell Biology, Domagkstrasse 3A, Muenster, D-48149 Germany

Lisa Schmidt, Sabine Przibilla, Gert von Bally, Björn Kemper

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

Jürgen Schnekenburger

University of Muenster, Department of Medicine B, Gastroenterological Molecular Cell Biology, Domagkstrasse 3A, Muenster, D-48149 Germany

J. Biomed. Opt. 15(4), 041509 (July 07, 2010). doi:10.1117/1.3449567
History: Received October 12, 2009; Revised March 19, 2010; Accepted March 29, 2010; Published July 07, 2010; Online July 07, 2010
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The understanding of biological reactions and evaluation of the significance for living cells strongly depends on the ability to visualize and quantify these processes. Digital holographic microscopy (DHM) enables quantitative phase contrast imaging for high resolution and minimal invasive live cell analysis without the need of labeling or complex sample preparation. However, due to the rather homogeneous intracellular refractive index, the phase contrast of subcellular structures is limited and often low. We analyze the impact of the specific manipulation of the intracellular refractive index by microinjection on the DHM phase contrast. Glycerol is chosen as osmolyte, which combines high solubility in aqueous solutions and biological compatibility. We show that the intracellular injection of glycerol causes a contrast enhancement that can be explained by a decrease of the cytosolic refractive index due to a water influx. The underlying principle is proven by experiments inducing cell shrinkage and with fixated cells. The integrity of the cell membrane is considered as a prerequisite and allows a reversible cell swelling and shrinking within a certain limit. The presented approach to control the intracellular phase contrast demonstrated for the example of DHM opens prospects for applications with other quantitative phase contrast imaging methods.

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© 2010 Society of Photo-Optical Instrumentation Engineers

Citation

Christina E. Rommel ; Christian Dierker ; Lisa Schmidt ; Sabine Przibilla ; Gert von Bally, et al.
"Contrast-enhanced digital holographic imaging of cellular structures by manipulating the intracellular refractive index", J. Biomed. Opt. 15(4), 041509 (July 07, 2010). ; http://dx.doi.org/10.1117/1.3449567


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