Microscopie holographique

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Microscopie holographique à contraste de phase
quantitative dans les cellules vivantes P.
Marquet1, E. Cuche2, J.-Y. Chatton1, C.
Depeursinge2, P. J. Magistretti1 1Institut de
physiologie, faculté de médecine, Université de
Lausanne, CH-1005 Lausanne, Switzerland 2Instit
ut dOptique appliquée, EPFL, CH-1015 Lausanne
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Electromagnetic waves
d
Mathematical expression
? -gt ?(d, n, ?)
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Light coherence
A) coherent source
d
B) non coherent source
random phase
d
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Sample illumination
Dimension detected IntensityJ/Sm2
(Intensity contrast microscopy)
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Origin of the Phase Shift
? --gt ?(d, next, nint)
nextlt nint
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Youngs interference
Phase and amplitude are recorded
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Principe of phase recording
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hologram
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Hologram reconstruction
Digital reconstruction
Numerical simulation of reference wave
diffraction by the hologram 1) intensity
contrast images 2) Quantitative phase contrast
images
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Holography set up
Characteristics Lateral resolution ? 400 nm
(0.75?/NA) z-Axis resolution ? 1..10 nm
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Applications
3D morphology and volume measurements in living
cells
Astrocyte in culture
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