An approach is proposed for removing the wave front curvature introduced by the microscope imaging objective in digital holography, which otherwise hinders the phase contrast imaging at reconstruction planes. The unwanted curvature is compensated by evaluating a correcting wave front at the hologram plane with no need for knowledge of the optical parameters, focal length of the imaging lens, or distances in the setup. Most importantly it is shown that a correction effect can be obtained at all reconstruction planes. Three different methods have been applied to evaluate the correction wave front and the methods are discussed in detail. The proposed approach is demonstrated by applying digital holography as a method of coherent microscopy for imaging amplitude and phase contrast of microstructures.
In microscopy, high magnifications are achievable for investigating micro-objects but the paradigm is that higher is the required magnification, lower is the depth of focus. For an object having a three-dimensional (3D) complex shape only a portion of it appears in good focus to the observer who is essentially looking at a single image plane. Actually, two approaches exist to obtain an extended focused image, both having severe limitations since the first requires mechanical scanning while the other one requires specially designed optics. We demonstrate that an extended focused image of an object can be obtained through digital holography without any mechanical scanning or special optical components. The conceptual novelty of the proposed approach lies in the fact that it is possible to completely exploit the unique feature of DH in extracting all the information content stored in hologram, amplitude and phase, to extend the depth of focus.
In this paper, we have investigated on the potentialities of digital holography for whole reconstruction of wavefields. We show that this technique can be efficiently used for obtaining quantitative information from the intensity and the phase distributions of the reconstructed field at different locations along the propagation direction. The basic concept and procedure of wavefield reconstruction for digital in-line holography is discussed. Numerical reconstructions of the wavefield from digitally recorded in-line hologram patterns and from simulated test patterns are presented. The potential of the method for analysing aberrated wave front has been exploited by applying the reconstruction procedure to astigmatic hologram patterns.
Aberrations and the distortions due to the imaging optics can be compensated in quantitative phase microscopy of thin phase objects by digital holography using a single hologram. The reconstructed quantitative phase microscopy phase distribution map can be directly corrected in the reconstructed image plane by a numerical method. To remove this unwanted aberration, in the special case of thin objects, the authors perform a two-dimensional fit with the Zernike polynomials of the reconstructed unwrapped phase. Subtraction of the fitted polynomial from the original phase map gives quantitative phase microscopy phase map free of aberrations.
9 February 2004 in Measurement Science and Technology
The unique deformability and the compliance ability of thin sheets on soft substrates attract much interest for studying the phenomena related to elastic instabilities as well as for sensing very weak forces such as those generated by live cells in vitro. However, the techniques used currently for producing such platforms are affected by a high degree of complexity and poor repeatability. Moreover, their deformability is usually used as a passive response to the action of an external force. Herein we propose a novel concept for a reliable and dynamic skin-over-liquid system made of a periodic array of highly compliant microbumps actuated through electrode-free electrohydrodynamic (EHD) pressure. We demonstrate that these structures are highly repeatable and capable of swelling and deflating easily under a simple thermal stimulation driven by the pyroelectric effect, thus providing a challenging platform that can be actively controlled at the microscale. Furthermore, we show the proof of principle by swelling these microbumps for mechanically stimulating live cells in vitro, thus opening the route to more reliable and easy to accomplish assays in the field of mechanobiology.