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Journal of Biomedical Optics • Open Access

Correlation between off-axis illumination and apodized phase-contrast: two complementary microscopic phase-imaging modes
Author(s): Radek Pelc; Zdenek Hostounsky; Tatsuro Otaki

Paper Abstract

Microscopic images of biological phase specimens of various optical thickness, acquired under off-axis illumination and apodized/conventional phase-contrast are compared. The luminance profiles in appropriately filtered apodized phase-contrast images compare well with those in the original off-axis illumination images. The two unfiltered image types also yield similar results in terms of quasi-three-dimensional surface (pseudo-relief) rendering, and thus are comparable in terms of the information contents (optical thickness map). However, the overall visual impression is very different as the visual cues to depth structure are present in the off-axis illumination images only. The comparison demonstrated in the present paper was made possible owing to apodization, which substantially reduces the "halo"/shade-off artifacts in the phase-contrast images. The results imply the possibility of combining the off-axis illumination and apodized phase-contrast imaging to examine specimens of medium optical thickness, in which the phase visualization capability of the two imaging modes substantially overlaps (e.g., larger cells or cell clusters).

Paper Details

Date Published: 1 September 2008
PDF: 12 pages
J. Biomed. Opt. 13(5) 054067 doi: 10.1117/1.2966716
Published in: Journal of Biomedical Optics Volume 13, Issue 5
Show Author Affiliations
Radek Pelc, Academy of Sciences of the Czech Republic (Czech Republic)
Zdenek Hostounsky
Tatsuro Otaki, Nikon Corp. (Japan)

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