Digital holography applied to microscopy

Christian Depeursinge*, Etienne Cuche, Pierre Marquet, Tristan Colomb, Pia Dahlgren, Anca M. Marian, Frederic Montfort, Pierre Magistretti

*Corresponding author for this work

Research output: Contribution to journalConference articlepeer-review

17 Scopus citations

Abstract

We report on new developments of a technique, called Digital Holographic Microscopy (DHM), for the numerical reconstruction of digital holograms taken in microscopy, which allows simultaneous amplitude and quantitative phase contrast imaging. The reconstruction method computes the propagation of the complex optical wavefront diffracted by the object and is used to determine the refractive index and/or shape of the object with accuracies in the nanometer range along the optical axis. The method requires the acquisition of a single hologram. The technique comprises the recording of a digital hologram of the specimen by means of a standard CCD camera at the exit of a Mach-Zender or Michelson type interferometer. The quantitative nature of the reconstructed phase distribution has been demonstrated by an application to surface profilometry where step height differences of a few nanometers have been measured. Another application takes place in biology for transmission phase-contrast imaging of living cells in culture. The resolution for thickness measurements depends on the refractive index of the specimen and a resolution of approximately 30 nanometers in height, and about half of a micron in width, has been achieved for living neural cells in cultures by using a high numerical aperture.

Original languageEnglish (US)
Pages (from-to)30-34
Number of pages5
JournalProceedings of SPIE - The International Society for Optical Engineering
Volume4659
DOIs
StatePublished - Jan 1 2002
EventPractical Holography XVI and Holographic Materials VIII - San Jose, United States
Duration: Jan 21 2002Jan 23 2002

Keywords

  • Cell culture imaging
  • Microscopy, holography
  • Phase contrast
  • Surface metrology

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Condensed Matter Physics
  • Computer Science Applications
  • Applied Mathematics
  • Electrical and Electronic Engineering

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