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Journal of Biomedical Optics

Contrast-enhanced digital holographic imaging of cellular structures by manipulating the intracellular refractive index
Author(s): Christina E. Rommel; Christian Dierker; Lisa Schmidt; Sabine Przibilla; Gert von Bally; Björn Kemper; Jürgen Schnekenburger
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Paper Abstract

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.

Paper Details

Date Published: 1 July 2010
PDF: 10 pages
J. Biomed. Opt. 15(4) 041509 doi: 10.1117/1.3449567
Published in: Journal of Biomedical Optics Volume 15, Issue 4
Show Author Affiliations
Christina E. Rommel, Westfälische Wilhelms-Univ. Münster (Germany)
Christian Dierker, Westfälische Wilhelms-Univ Münster (Germany)
Lisa Schmidt, Westfälische Wilhelms-Univ. Münster (Germany)
Sabine Przibilla, Westfälische Wilhelms-Univ. Münster (Germany)
Gert von Bally, Westfälische Wilhelms-Univ. Münster (Germany)
Björn Kemper, Westfälische Wilhelms-Univ. Münster (Germany)
Jürgen Schnekenburger, Westfälische Wilhelms-Univ Münster (Germany)


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