Quantitative phase-contrast microscopy by angular spectrum digital holography

C. J. Mann, M. K. Kim

Research output: Chapter in Book/Report/Conference proceedingConference contribution

19 Scopus citations

Abstract

Techniques of digital holography are improved in order to obtain high-resolution, high-fidelity images of quantitative phase-contrast microscopy. In particular, the angular spectrum method of calculating the holographic optical field is seen to have several advantages over the more commonly used Fresnel transformation or Huygens convolution method. Spurious noise and interference components can be tightly controlled through the analysis and filtering of the angular spectrum. The reconstruction distance does not have a lower limit and the off-axis angle between the object and reference can be lower than the Fresnel requirement and still be able to cleanly separate out the zero-order background. Holographic phase images are largely immune from the coherent noise common in amplitude images. Together with the use of a miniature pulsed laser, the resulting images have 0.5 μm diffraction-limited lateral resolution and the phase profile is accurate to about 30 nm of optical path length. SKOV-3 (ovarian cancer cells) and HUVEC (human umbilical vein endothelial cells) are imaged that display intra-cellular and intra-nuclear organelles with clarity and quantitative accuracy. The technique clearly exceeds currently available methods in phase-contrast optical microscopy in the level of resolution and detail, and provides a new modality for imaging morphology of cellular and intracellular structures that is not currently available.

Original languageEnglish (US)
Title of host publicationProceedings of SPIE - The International Society for Optical Engineering
DOIs
StatePublished - May 8 2006
Externally publishedYes
EventThree-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIII - San Jose, CA, United States
Duration: Jan 24 2006Jan 26 2006

Publication series

NameProceedings of SPIE - The International Society for Optical Engineering
Volume6090
ISSN (Print)0277-786X

Other

OtherThree-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIII
CountryUnited States
CitySan Jose, CA
Period1/24/061/26/06

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Keywords

  • Angular spectrum
  • Digital holography
  • Phase microscopy

ASJC Scopus subject areas

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

Cite this

Mann, C. J., & Kim, M. K. (2006). Quantitative phase-contrast microscopy by angular spectrum digital holography. In Proceedings of SPIE - The International Society for Optical Engineering [60900B] (Proceedings of SPIE - The International Society for Optical Engineering; Vol. 6090). https://doi.org/10.1117/12.645412