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Optics Express

Optics Express

  • Editor: Micha
  • Vol. 13, Iss. 23 — Nov. 14, 2005
  • pp: 9361–9373

Measurement of the integral refractive index and dynamic cell morphometry of living cells with digital holographic microscopy

Benjamin Rappaz, Pierre Marquet, Etienne Cuche, Yves Emery, Christian Depeursinge, and Pierre J. Magistretti  »View Author Affiliations

Optics Express, Vol. 13, Issue 23, pp. 9361-9373 (2005)

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We have developed a digital holographic microscope (DHM), in a transmission mode, adapted to the quantitative study of cellular dynamics. Living cells in culture are optically probed by measuring the phase shift they produce on the transmitted wave front. The high temporal stability of the phase signal, equivalent to λ/1800, and the low acquisition time (∼20μs) enable to monitor cellular dynamics processes. An experimental procedure allowing to calculate both the integral refractive index and the cellular thickness (morphometry) from the measured phase shift is presented. Specifically, the method has been applied to study the dynamics of neurons in culture during a hypotonic stress. Such stress produces a paradoxical decrease of the phase which can be entirely resolved by applying the methodological approach described in this article; indeed the method allows to determine independently the thickness and the integral refractive index of cells.

© 2005 Optical Society of America

OCIS Codes
(090.1760) Holography : Computer holography
(120.3940) Instrumentation, measurement, and metrology : Metrology
(120.5050) Instrumentation, measurement, and metrology : Phase measurement
(170.1530) Medical optics and biotechnology : Cell analysis

ToC Category:
Research Papers

Original Manuscript: September 20, 2005
Revised Manuscript: October 31, 2005
Published: November 14, 2005

Benjamin Rappaz, Pierre Marquet, Etienne Cuche, Yves Emery, Christian Depeursinge, and Pierre Magistretti, "Measurement of the integral refractive index and dynamic cell morphometry of living cells with digital holographic microscopy," Opt. Express 13, 9361-9373 (2005)

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