Quantitative analysis of three-dimensional biological cells using interferometric microscopy

Natan T. Shaked, Adam Wax

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review

Abstract

Live biological cells are three-dimensional microscopic objects that constantly adjust their sizes, shapes and other biophysical features. Wide-field digital interferometry (WFDI) is a holographic technique that is able to record the complex wavefront of the light which has interacted with in-vitro cells in a single camera exposure, where no exogenous contrast agents are required. However, simple quasi-three-dimensional holographic visualization of the cell phase profiles need not be the end of the process. Quantitative analysis should permit extraction of numerical parameters which are useful for cytology or medical diagnosis. Using a transmission-mode setup, the phase profile represents the multiplication between the integral refractive index and the thickness of the sample. These coupled variables may not be distinct when acquiring the phase profiles of dynamic cells. Many morphological parameters which are useful for cell biologists are based on the cell thickness profile rather than on its phase profile. We first overview methods to decouple the cell thickness and its refractive index using the WFDI-based phase profile. Then, we present a whole-cell-imaging approach which is able to extract useful numerical parameters on the cells even in cases where decoupling of cell thickness and refractive index is not possible or desired.

Original languageEnglish
Title of host publicationThree-Dimensional Imaging, Visualization, and Display 2011
PublisherSPIE
ISBN (Print)9780819486172
DOIs
StatePublished - 2011
EventThree-Dimensional Imaging, Visualization, and Display 2011 - Orlando, FL, United States
Duration: 27 Apr 201128 Apr 2011

Publication series

NameProceedings of SPIE - The International Society for Optical Engineering
Volume8043
ISSN (Print)0277-786X
ISSN (Electronic)1996-756X

Conference

ConferenceThree-Dimensional Imaging, Visualization, and Display 2011
Country/TerritoryUnited States
CityOrlando, FL
Period27/04/1128/04/11

Keywords

  • Cell analysis
  • Interference microscopy
  • Phase holography

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