Diffractive Optical Elements Investigation in the Phase Domain

In this work we demonstrate the advantages of investigating diffractive optical elements in the phase domain. In this regime we can detect features that are not restrained by the diffraction limit and relate them to the geometrical and optical properties of the sample under test. To accomplish that, we use the custom made spectral high resolution interference microscope. Phase map recordings allow for easier and more precise localization of the positions, where phase changes happen. We show the localization capabilities by detecting phase singularities created by a trench. We also apply the concept to abrupt phase jumps of a phase diffractive component and determine the achievable resolution.

Numerical reconstruction of digital holograms

Etienne Cuche

This thesis presents a new method for the numerical reconstruction of digital holograms recorded in the off-axis configuration using a CCD camera. The propagation of the reconstructed wave front, from the hologram plane to an observation plane, is achieved by a calculation of scalar diffraction in the Fresnel approximation. An image representing the distribution of intensity at the surface of the object can be obtained by calculating the intensity of the reconstructed wave front. The focusing of this image is performed digitally by adjusting a parameter which defines the distance between the hologram and the observation plane. A second image representing the phase distribution at the surface of the object can be obtained from the same hologram by calculating the argument of the reconstructed wave front. We show here that for getting a phase distribution that represents only the contribution of the object wave, the hologram or the reconstructed wave front must be multiplied by a computed replica of the complex conjugate of the wave used as reference during the recording. For a plane reference wave, the reconstruction of the phase distribution requires a precise adjustment of two digital parameters defining its propagation direction. An application to surface profilometry has demonstrated the quantitative nature of the obtained phase distribution which gives access to the topography of the object. A vertical resolution of λ/100 has been estimated on the basis of a preliminary study. The introduction of a microscope objective along the optical path of the object wave enhances the transverse resolution of the method. In this case, the wave front deformation produced by the microscope objective can be digitally compensated. Under the assumption of a paraboloidal deformation, we show that the wave front curvature can be numerically corrected by multiplication with an array of complex numbers computed using the complex conjugate of the analytical expression describing the defocusing aberration. A preliminary study indicates that the transverse resolution of the method approaches that of classical optical microscopy (diffraction limit). The application of a digital technique of spatial filtering allows for the elimination of the zero order of diffraction and of the twin image. This operation enhances the signal to background ratio. We show also that the influence of diffraction effects associated to the numerical reconstruction method can be reduced by apodization of the hologram aperture. This operation is performed digitally by means of a procedure which defines the transmission of the hologram aperture using a cubic spline interpolation.

EPFL2000

Point spread function models for digital holographic microscopy

Christian Depeursinge, Mihaela Marian

Few models, based on the diffraction theory, are proposed in order to evaluate the point spread function of different microscope objectives used in a digital holographic microscope. Because in holography the phase information is essential, a 3D amplitude point spread function (APSF), modulus and phase, is necessary, in order to properly deconvolute the 3D images obtained. Scalar Debye theory, paraxial approximation and vectorial Debye theory are used to solve the diffraction problem and the theoretical predicted 3D APSFs obtained. with these models are compared.

SPIE2004

Resolution and Complex Deconvolution in Holographic Microscopy

Yann Jérôme Michel Pascal Cotte

Interest in high resolution imaging techniques has recently multiplied due to their importance in bio-medical research. Quantitative phase measurements by holographic microscopy is an extraordinary tool to gain new understanding of transparent biological samples. The extremely high demand on resolution well beyond the diffraction limit, however, sets new benchmarks for imaging techniques. Digital Holographic Microscopy is an interferometric method providing access to the complex wave front. Its capacity to image amplitude and quantitative phase simultaneously makes it an attractive research tool in many fields of biological research. It is on the verge of becoming an appealing alternative to classical fluorescence microscopy. For intensity-based microscopy, however, super-resolution methods are well established. In this thesis, approaches to unleash resolution for phase imaging techniques are explored. Based on truncated inverse filtering, a theory for deconvolution of complex fields is developed. It is a post-processing method that does not require any additional optics in the holographic microscopy setup. Gain in resolution arises by accessing the object's complex field – containing the information encoded in the phase – and deconvolving it with the characteristic coherent transfer function (CTF). By efficient harvest of a diffraction limited bandpass, complex deconvolution is demonstrated to exceed the coherent resolution limit. A novel method, called a complex point source, serves to characterize the holographic microscopy system. It consists of a coherently illuminated nano-metric hole, located on a conventional microscope slide, most common in bio-microscopy. A thin opaque film is directly evaporated on the slide and the circular sub-wavelength structures are drilled with a focused ion beam. Theoretical as well as practical work on transmission properties of apertures confirms the advantageous signal-to-noise yield of proposed method. More abstractly, the thus gained experimental amplitude point spread function is demonstrated to be suitable for experimental CTF reconstruction. Herefrom, experimental parameters are extracted and introduced into scalar and vectorial Debye theory, called synthetic CTF, adaptable to realistic imaging conditions. Also, it allows to study the role of noise in the context of complex field deconvolution. The resolution power of holographic microscopy systems is systematically examined with pairs of nano-metric apertures separated by sub-resolution pitches. Theory of information content is elaborated depending on different experimental configurations. Resolution extends beyond classical Abbe's limit by introducing spatially a varying phase into the illumination beam of a phase imaging system. In a last step, the novel deconvolution method is generalized to three-dimensional image processing of complex fields. By combining scattering theory and signal processing, the method is demonstrated to yield the reconstruction of the scattering object field under non-design microscope objective imaging conditions. The suggested technique is best suited for an implementation in high-resolution diffraction tomography based on sample and/or illumination rotation.