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  4. Quantitative Measurement of absolute cell volume and intracellular integral refractive index (RI) with Dual-wavelength Digital Holographic Microscopy (DHM)
 
conference paper

Quantitative Measurement of absolute cell volume and intracellular integral refractive index (RI) with Dual-wavelength Digital Holographic Microscopy (DHM)

Boss, Daniel  
•
Kuehn, Jonas
•
Depeursinge, Christian  
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2012
Biophotonics: Photonic Solutions For Better Health Care Iii
Conference on Biophotonics - Photonic Solutions for Better Health Care III

Quantitative Phase Imaging techniques including DHM have been applied recently in the field of cell imaging to monitor and quantify non-invasively dynamic cellular processes modifying cell morphology and/or content. Concretely, the DHM phase signal is highly sensitive to cell thickness and intracellular integral RI variations associated with transmembrane water movements. As net water flow across the cell membrane leads at the same time to changes in cell thickness and intracellular RI, the interpretation of phase signal variations remains difficult. To overcome this drawback, we have developed a Dual-wavelength Digital Holographic Microscopy (DHM) setup allowing to separately measure, with a single CCD camera acquisition, thickness and integral RI of living cells. The method is based on the use of an absorbing dye that enhances the refractive index dispersion of the extracellular medium. Practically, two significantly different phase signals can be obtained when measuring at two appropriate wavelengths. From the two phase measurements, both cell RI and thickness can be univocally determined.

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Type
conference paper
DOI
10.1117/12.922598
Web of Science ID

WOS:000305701200006

Author(s)
Boss, Daniel  
Kuehn, Jonas
Depeursinge, Christian  
Magistretti, Pierre J.  
Marquet, Pierre
Date Issued

2012

Publisher

Spie-Int Soc Optical Engineering, Po Box 10, Bellingham, Wa 98227-0010 Usa

Published in
Biophotonics: Photonic Solutions For Better Health Care Iii
ISBN of the book

978-0-8194-9119-0

Series title/Series vol.

Proceedings of SPIE; 8427

Start page

84270B

Subjects

Digital Holographic Microscopy

•

Absolute cell volume

•

Phase decoupling procedure

•

Water membrane permeability

•

Dry mass concentration

•

Living Cells

•

Morphometry

Editorial or Peer reviewed

NON-REVIEWED

Written at

EPFL

EPFL units
LOA  
LNDC  
Event nameEvent placeEvent date
Conference on Biophotonics - Photonic Solutions for Better Health Care III

Brussels, BELGIUM

Apr 16-19, 2012

Available on Infoscience
July 20, 2012
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/84047
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