Abstract
Differential image contrast (DIC), through the numerical managing and manipulation of complex wavefronts obtained by digital holography (DH), is investigated. We name the approach Dynamical Differential Holographic Image Contrast (DDHIC). DDHIC dispenses from special optics and/or complex setup configurations with moveable components, as usually occurs in classical DIC, that is not well-suited for investigating objects experiencing dynamic evolution during the measurement. In fact, the technique presented here, is useful for floating samples since it allows, from a single recording, to set a posteriori the best conditions for DIC imaging in conjunction with the numerical focusing feature of DH. By DDHIC, the movies can be easily built-up to offering dynamic representation of phase-contrast along all directions, thus improving the visualization. Furthermore, the dynamic representation is useful for making the proper choice of other key parameters of DIC such as the amount of shear and the bias, with the aim to optimize the visualized phase-contrast imaging as favorite representation for bio-scientists. Investigation is performed on various biological samples.
Highlights
Behavior and growth of cells and microorganisms is a key issue in biology and biomedical research fields
In this way a novel concept of Differential image contrast (DIC) in microscopy can be envisaged by using digital holography (DH), we named here DDHIC (Dynamical Differential Holographic Image Contrast) where the numerical implementation of DIC images are obtained manipulating numerically the DH phase-maps
DDHIC images for different values of the bias are displayed in Figs. 6(a)–6(i), where the shear direction is kept fixed at 45° and the shear quantity, selected before, at 4RP
Summary
Behavior and growth of cells and microorganisms is a key issue in biology and biomedical research fields. The flexibility of DH is employed to perform quantitative PCI mapping as well as synthetic DIC imaging of biological sample. Dynamic display of DIC phase-contrast images along all directions can enhance the visualization of all details of the object under investigation. In this way a novel concept of DIC in microscopy can be envisaged by using DH, we named here DDHIC (Dynamical Differential Holographic Image Contrast) where the numerical implementation of DIC images are obtained manipulating numerically the DH phase-maps. We illustrate the novel concept of DDHIC, describing the procedure, to follow step by step with the aim to optimize the phase contrast demonstrating its valuable effect in visualizing static and dynamic biological samples
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