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Quantitative Phase Contrast Microscopy of Living Astrocytes By Numerical Reconstruction of Digital Holograms

Published online by Cambridge University Press:  02 July 2020

P. Marquet ,
E. Cuche ,
J.-Y. Chatton ,
Ch. Depeursinge  and
P.J. Magistretti
P. Marquet
Affiliation:
institute of Physiology and Department of Neurology, University of Lausanne, CH- 1005 Lausanne, Switzerland
E. Cuche
Affiliation:
Applied Optics Institute, Swiss Federal institute of Technology, CH-1005 Lausanne, Switzerland
J.-Y. Chatton
Affiliation:
institute of Physiology and Department of Neurology, University of Lausanne, CH- 1005 Lausanne, Switzerland
Ch. Depeursinge
Affiliation:
Applied Optics Institute, Swiss Federal institute of Technology, CH-1005 Lausanne, Switzerland
P.J. Magistretti
Affiliation:
institute of Physiology and Department of Neurology, University of Lausanne, CH- 1005 Lausanne, Switzerland
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Extract

We have developed a method of numerical holography which allows simultaneous intensity and quantitative phase contrast imaging. Measurement of optical path length provides precise information about the cell morphology. As presented in Fig. 1, the experimental set-up for the recording of digital holograms is basically a Mach-Zender interferometer. At the exit of the interferometer, a CCD camera records a hologram which is made by the interference between a reference wave R and the object wave O that is transmitted by the biological sample. In order to obtain a plane wave as reference, a beam expander (B.E.) including a pinhole is introduced in the reference arm and enlarges the beam diameter to about 2cm. A similar system which produces a smaller beam diameter (≈5mm) is inserted in the objet arm. To perform holography in the off-axis geometry, the mirror in the reference arm is oriented in such a way that the reference wave reaches the CCD camera with a small incidence angle.

Type
Novel Approaches to Microscopy Of Living Cells
Information
Microscopy and Microanalysis , Volume 5 , Issue S2: Proceedings: Microscopy & Microanalysis '99, Microscopy Society of America 57th Annual Meeting, Microbeam Analysis Society 33rd Annual Meeting, Portland, Oregon August 1-5, 1999 , August 1999 , pp. 1054 - 1055
Copyright
Copyright © Microscopy Society of America

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References

1)Cuche, E. etal.,Opt. Lett., in press (1999)Google Scholar