Numerical aberrations compensation and polarization imaging in digital holographic microscopy

In this thesis, we describe a method for the numerical reconstruction of the complete wavefront properties from a single digital hologram: the amplitude, the phase and the polarization state. For this purpose, we present the principle of digital holographic microscopy (DHM) and the numerical reconstruction process which consists of propagating numerically a wavefront from the hologram plane to the reconstruction plane. We then define the different parameters of a Numerical Parametric Lens (NPL) introduced in the reconstruction plane that should be precisely adjusted to achieve a correct reconstruction. We demonstrate that automatic procedures not only allow to adjust these parameters, but in addition, to completely compensate for the phase aberrations. The method consists in computing directly from the hologram a NPL defined by standard or Zernike polynomials without prior knowledge of physical setup values (microscope objective focal length, distance between the object and the objective...). This method enables to reconstruct correct and accurate phase distributions, even in the presence of strong and high order aberrations. Furthermore, we show that this method allows to compensate for the curvature of specimen. The NPL parameters obtained by Zernike polynomial fit give quantitative measurements of micro-optics aberrations and the reconstructed images reveal their surface defects and roughness. Examples with micro-lenses and a metallic sphere are presented. Then, this NPL is introduced in the hologram plane and allows, as a system of optical lenses, numerical magnification, complete aberration compensation in DHM (correction of image distortions and phase aberrations) and shifting. This NPL can be automatically computed by polynomial fit, but it can also be defined by a calibration method called Reference Conjugated Hologram (RCH). We demonstrate the power of the method by the reconstruction of non-aberrated wavefronts from holograms recorded specifically with high orders aberrations introduced by a tilted thick plate, or by a cylindrical lens or by a lens ball used instead of the microscope objective. Finally, we present a modified digital holographic microscope permitting the reconstruction of the polarization state of a wavefront. The principle consists in using two reference waves polarized orthogonally that interfere with an object wave. Then, the two wavefronts are reconstructed separately from the same hologram and are processed to image the polarization state in terms of Jones vector components. Simulated and experimental data are compared to a theoretical model in order to evaluate the precision limit of the method for different polarization states of the object wave. We apply this technique to image the birefringence and the dichroism induced in a stressed polymethylmethacrylate sample (PMMA), in a bent optical fiber and in a thin concrete specimen. To evaluate the precision of the phase difference measurement in DHM design, the birefringence induced by internal stress in an optical fiber is measured and compared to the birefringence profile captured by a standard method, which had been developed to obtain high-resolution birefringence profiles of optical fibers. A 6 degrees phase difference resolution is obtained, comparable with standard imaging polariscope, but with the advantage of a single acquisition allowing real-time reconstruction

3D perception of numerical hologram reconstructions enhanced by motion and stereo

We investigated the question of how the perception of 3D information of digital holograms reconstructed numerically and presented on conventional displays depends on motion and stereoscopic presentation. Perceived depth in an adjustable random pattern stereogram was matched to the depth in holographic objects. The objects in holograms were a microscopic biological cell and a macroscopic coil. Stereoscopic presentation increased perceived depth substantially in comparison to non-stereoscopic presentation. When stereoscopic cues were weak or absent e.g. because of blur, motion increased perceived depth considerably. However, when stereoscopic cues were strong, the effect of motion was small. In conclusion, for the maximisation of perceived 3D information of holograms on conventional displays, it seems highly beneficial to use the combination of motion and stereoscopic presentation

Nanoindentation and birefringence measurements on fused silica specimen exposed to low-energy femtosecond pulses,” Opt.

Abstract: Femtosecond laser pulses used in a regime below the ablation threshold have two noticeable effects on Fused Silica (a-SiO2): they locally increase the material refractive index and modify its HF etching selectivity. The nature of the structural changes induced by femtosecond laser pulses in fused silica is not fully understood. In this paper, we report on nanoindentation and birefringence measurements on fused silica exposed to low-energy femtosecond laser pulses. Our findings further back the hypothesis of localized densification effect even at low energy regime

Living specimen tomography by digital holographic microscopy: morphometry of testate amoeba

This paper presents an optical diffraction tomography technique based on digital holographic microscopy. Quantitative 2-dimensional phase images are acquired for regularly-spaced angular positions of the specimen covering a total angle of π, allowing to built 3-dimensional quantitative refractive index distributions by an inverse Radon transform. A 20x magnification allows a resolution better than 3 μm in all three dimensions, with accuracy better than 0.01 for the refractive index measurements. This technique is for the first time to our knowledge applied to living specimen (testate amoeba, Protista). Morphometric measurements are extracted from the tomographic reconstructions, showing that the commonly used method for testate amoeba biovolume evaluation leads to systematic under evaluations by about 50%

Versatile spectral modulation of a broadband source for digital holographic microscopy

We demonstrate the potential of spatial light modulators for the spectral control of a broadband source in digital holographic microscopy. Used in a ‘pulse-shaping’ geometry, the spatial light modulator provides a versatile control over the bandwidth and wavelength of the light source. The control of these properties enables adaptation to various experimental conditions. As a first application, we show that the source bandwidth can be adapted to the off-axis geometry to provide quantitative phase imaging over the whole field of view. As a second application, we generate sequences of appropriate wavelengths for a hierarchical optical phase unwrapping algorithm, which enables the measurement of the topography of high-aspect ratio structures without phase ambiguity. Examples are given with step heights up to 50 um

Several micron-range measurements with sub-nanometric resolution by the use of dual-wavelength digital holography and vertical scanning

Reflection digital holographic microscopy (DHM) is a very powerful technique allowing measuring topography with a sub-nanometer axial resolution from a single hologram acquisition. But as most of interferometer methods, the vertical range is limited to half the wavelength if numerical unwrapping procedure could not be applied (very high aspect ratio specimen). Nevertheless, it was already demonstrated that the use of dual-wavelength DHM allows increasing the vertical range up to several microns by saving the single wavelength resolution if conditions about phase noise are fulfilled (the higher the synthetic wavelength, the smaller the phase noise has to be). In this paper, we will demonstrate that the choice of a synthetic wavelength of about 17 microns allows measuring precisely a 4.463μm certified step. Furthermore, we will show the feasibility of a sub-nanometer resolution on a range higher than the synthetic wavelength by being able to map the dual- wavelength measurement on data acquired from a vertical scanning process, which precision is about 1 μm