Using a digital holographic microscope setup, it is possible to measure dynamic volume changes in living cells. The cells were investigated time-dependently in transmission mode for different kinds of stimuli affecting their morphology. The measured phase shift was correlated to the cellular optical thickness, and then of the cell volume as well as the refractive index were calculated and interpreted. For the characterization of the digital holographic microscope setup, we have developed a transparent three-dimensional (3-D) reference chart that can be used as a lateral resolution chart and step-height resolution chart included in one substrate. For the monitoring of living cells, a biocompatible and autoclavable flow chamber was designed, which allows us to add, exchange, or dilute the fluid within the flow chamber. An integrated changeable coverslip enables inverse microscopic applications. Trypsinization, cell swelling and shrinking induced by osmolarity changes, and apoptosis served as model processes to elucidate the potential of the digital holographic microscopy (DHM).
A novel implementation of lensless multiwavelength digital holography with autocalibration of temporal phase shifts and artificial wavelength is presented. The algorithm we used to calculate the phase shifts was previously proposed [Opt. Lett.29 183 (2004)] and, to our knowledge, is now used for the first time in lensless holography. Because precise knowledge of the generated artificial wavelength is crucial for absolute measurement accuracy, a simple and efficient method to determine the artificial wavelength directly is presented. The calibration method is based on a simple modification of the experimental setup and needs just one additional image acquisition per wavelength. The results of shape measurement of a metallic test object with a rough surface and steep edges are shown and the measurement accuracy is discussed.
We report on an experimental realization of polarization-holographic optical elements (PHOEs) for laser beam shaping. The PHOEs were written into an azobenzene polymer layer by illumination with linearly polarized, focused light with spatially varying orientation. We propose a noniterative design method for the calculation of PHOEs that transform laser beams into arbitrary rotationally symmetric or separable intensity distributions such as flattop or ring profiles. The experimentally observed intensity distributions are compared with those predicted by numerical simulations based on the Fresnel diffraction approximation. Diffraction efficiencies up to 79% were determined experimentally.
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