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  4. Sub-cellular quantitative optical diffraction tomography with digital holographic microscopy
 
conference paper

Sub-cellular quantitative optical diffraction tomography with digital holographic microscopy

Charrière, F.  
•
Kühn, J.
•
Colomb, T.  
Show more
2007
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V
Photonics West

Digital holographic microscopy (DHM) is an interferometric technique, providing quantitative mapping of the phase shift induced by semi- transparent microscopic specimens, such as cells, with sub-wavelenght resolution along the optical axis. Thanks to actual PCs and CCDs, DHM provides nowadays cost-effective instruments for real-time measurements at very high acquisition rates, with sub-micron transverse resolution. However, DHM phase images do not reveal the three-dimensional (3D) internal distribution of refractive index, but a phase shift resulting from a mean refractive index (RI) integrated over the cellular thickness. Standard optical diffraction tomography (ODT) techniques can be efficiently applied to reveal internal structures and to measure 3D RI spatial distributions, by recording 2D DHM phase data for different sample orientations or illumination beam direction, in order to fill up entirely the Ewald sphere in the Fourier space. The 3D refractive index can then be reconstructed, even in the direct space with backpropagation algorithms or from the Fourier space with inverse Fourier transform. The presented technique opens wide perspectives in 3D cell imaging: the DHM-based micro-tomography furnishes invaluable data on the cell components optical properties, potentially leading to information about organelles intracellular distribution. Results obtained on biological specimens will be presented. Morphometric measurements can be extracted from the tomographic data, by detection based on the refractive index contrast within the 3D reconstructions. Results and perspectives about sub- cellular organelles identification inside the cell will also be exposed.

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Type
conference paper
DOI
10.1117/12.700540
Web of Science ID

WOS:000245855200016

Author(s)
Charrière, F.  
Kühn, J.
Colomb, T.  
Cuche, E.  
Marquet, P.
Depeursinge, C.  
Date Issued

2007

Publisher

SPIE

Published in
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V
ISBN of the book

0819465542

Volume

6441

Subjects

[MVD]

•

3D imaging

•

Cell imaging

•

Digital holography

•

Tomography

URL

URL

http://spie.org/x648.html?product_id=700540
Editorial or Peer reviewed

NON-REVIEWED

Written at

EPFL

EPFL units
LOA  
Event nameEvent placeEvent date
Photonics West

San Jose, CA

January 20-25, 2007

Available on Infoscience
July 20, 2009
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/41627
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