We propose and test a focus plane determination method that computes the digital refocus distance of an object investigated by digital holographic microscopy working in transmission. For this purpose we analyze the integrated amplitude modulus as a function of the digital holographic reconstruction distance. It is shown that when the focus distance is reached, the integrated amplitude is minimum for pure amplitude object and maximum for pure phase object. After a theoretical analysis, the method is demonstrated on actual digital holograms for the refocusing of pure amplitude and of pure phase microscopic samples.
The dynamics of a vesicle suspension in a shear flow between parallel plates has been investigated under microgravity conditions, where vesicles are only submitted to hydrodynamic effects such as lift forces due to the presence of walls and drag forces. The temporal evolution of the spatial distribution of the vesicles has been recorded thanks to digital holographic microscopy, during parabolic flights and under normal gravity conditions. The collected data demonstrates that vesicles are pushed away from the walls with a lift velocity proportional toγR 3 /z 2 whereγ is the shear rate, R the vesicle radius and z its distance from the wall. This scaling as well as the dependence of the lift velocity upon vesicle aspect ratio are consistent with theoretical predictions by Olla [J. Phys. II France 7, 1533-1540(1997].
We investigate the use of a digital holographic microscope working in partially coherent illumination to study in three dimensions a micrometer-size particle flow. The phenomenon under investigation rapidly varies in such a way that it is necessary to record, for every camera frame, the complete holographic information for further processing. For this purpose, we implement the Fourier-transform method for optical amplitude extraction. The suspension of particles is flowing in a split-flow lateral-transport thin separation cell that is usually used to separate the species by their sizes. Details of the optical implementation are provided. Examples of reconstructed images of different particle sizes are shown, and a particle-velocity measurement technique that is based on the blurred holographic image is exploited.
A mini splitterless-split-flow thin fractionation (SPLITT) device has been developed to achieve fast separations of micrometer-sized species. In this device, inlet and outlet steps have replaced the splitters, which are common to conventional SPLITT channels. By elimination of the splitters, it becomes straightforward to reduce channel dimensions while maintaining the classic method of fabrication. Reduced dimension channels allow high axial velocity at relatively low flow rate. These high axial velocities generate an enhancement of inertial lift forces and hydrodynamic shear-induced diffusion. Experiments carried out with particulate and biological species in a mini step-SPLITT channel demonstrate that these hydrodynamic effects yield highly enriched fractions of smaller species from binary mixtures.
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