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8 February 2011 Manipulating intracellular refractive index for contrast-enhanced digital holographic imaging of subcellular structures
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The online analysis of rapid cellular processes by morphological alterations strongly depends on the ability to rapidly visualize and to quantify cell shape and intracellular structures. 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 homogenous intracellular refractive index, the phase contrast of subcellular structures is limited and often low. We analyzed the impact of specific intracellular refractive index manipulation by microinjection of refractive index changing agents on the DHM phase contrast. Glycerol was chosen as osmolyte, which combines high solubility in aqueous solutions and cellular compatibility. We present data showing 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 was proven by experiments inducing cell shrinkage and protein concentration. The integrity of cell membranes 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 DHM opens also prospects for application with other quantitative phase contrast imaging technologies.
© (2011) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Christina E. Rommel, Christian Dierker, Lisa Schmidt, Sabine Przibilla, Gert von Bally, Björn Kemper, and Jürgen Schnekenburger "Manipulating intracellular refractive index for contrast-enhanced digital holographic imaging of subcellular structures", Proc. SPIE 7902, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues IX, 790203 (8 February 2011);

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