SummaryWe propose an extension to Nomarski differential interference contrast microscopy that enables isotropic linear phase imaging. The method combines phase shifting, two directions of shear and Fourier-space integration using a modified spiral phase transform. We simulated the method using a phantom object with spatially varying amplitude and phase. Simulated results show good agreement between the final phase image and the object phase, and demonstrate resistance to imaging noise.

SummaryDifferential interference contrast (DIC) is frequently used in conventional 2D biological microscopy. Our recent investigations into producing a 3D DIC microscope (in both conventional and confocal modes) have uncovered a fundamental difficulty: namely that the phase gradient images of DIC microscopy cannot be visualized using standard digital image processing and reconstruction techniques, as commonly used elsewhere in microscopy. We discuss two approaches to the problem of preparing gradient images for 3D visualization: integration and the Hilbert transform. After applying the Hilbert transform, the dataset can then be visualized in 3D using standard techniques. We find that the Hilbert transform provides a rapid qualitative pre-processing technique for 3D visualization for a wide range of biological specimens in DIC microscopy, including chromosomes, which we use in this study.

This study examined perceptual differentiation of specular from diffuse shading for the recovery of surface color and gloss. In Experiment 1, we parametrically varied the mesoscale relief height of globally planar surfaces, specular sharpness and the orientation of the surface relative to the light source. We obtained psychophysical matches for perceived color saturation and value (HSV), but also considered whether the main effects could be influenced by color space used when transforming data to perceptually-uniform CIE LCH space. Results revealed strong interactions between perceived color attributes and the lighting conditions, the structure of specular reflections, and surface relief. Declines in saturation were observed with increasing specular roughness (using an HSV color representation), but no similar decline was observed in chroma (using a CIE LCH color representation). Experiment 2 found strong negative correlations between perceived gloss and specular roughness. Perceived gloss also depended on mesoscopic relief height and orientation of the surface relative to the light source. Declines in perceived gloss moderately accounted for the variability in color saturation and value matches obtained in Experiment 1. We found information about perceived specular coverage could further improve the model's accountability of perceived color saturation and lightness (Experiment 3). These findings together suggest that perceived color saturation and color value depends on the visual system's ability to distinguish the underlying diffuse shading from specular highlights in images.

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