Research Papers: Imaging

Measurement of absolute cell volume, osmotic membrane water permeability, and refractive index of transmembrane water and solute flux by digital holographic microscopy

[+] Author Affiliations
Daniel Boss, Pascal Jourdain, Pierre J. Magistretti, Pierre Marquet

Ecole Polytechnique Fédérale de Lausanne, Laboratory of Neuroenergetics and Cellular Dynamics, Lausanne, Switzerland

Jonas Kühn

Centre de Neurosciences Psychiatriques, Département de Psychiatrie DP-CHUV, Prilly-Lausanne, Switzerland

Christian Depeursinge

Ecole Polytechnique Fédérale de Lausanne, Microvision and Microdiagnostics Group, Lausanne, Switzerland

J. Biomed. Opt. 18(3), 036007 (Mar 13, 2013). doi:10.1117/1.JBO.18.3.036007
History: Received October 4, 2012; Revised December 9, 2012; Accepted January 2, 2013
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Abstract.  A dual-wavelength digital holographic microscope to measure absolute volume of living cells is proposed. The optical setup allows us to reconstruct two quantitative phase contrast images at two different wavelengths from a single hologram acquisition. When adding the absorbing dye fast green FCF as a dispersive agent to the extracellular medium, cellular thickness can be univocally determined in the full field of view. In addition to the absolute cell volume, the method can be applied to derive important biophysical parameters of living cells including osmotic membrane water permeability coefficient and the integral intracellular refractive index (RI). Further, the RI of transmembrane flux can be determined giving an indication about the nature of transported solutes. The proposed method is applied to cultured human embryonic kidney cells, Chinese hamster ovary cells, human red blood cells, mouse cortical astrocytes, and neurons.

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© 2013 Society of Photo-Optical Instrumentation Engineers

Citation

Daniel Boss ; Jonas Kühn ; Pascal Jourdain ; Christian Depeursinge ; Pierre J. Magistretti, et al.
"Measurement of absolute cell volume, osmotic membrane water permeability, and refractive index of transmembrane water and solute flux by digital holographic microscopy", J. Biomed. Opt. 18(3), 036007 (Mar 13, 2013). ; http://dx.doi.org/10.1117/1.JBO.18.3.036007


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