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Proceedings Paper

The impact of the depth of field on cytogenetic image quality in scanning microscopy
Author(s): Yuchen Qiu; Xiaodong Chen; Yuhua Li; Bin Zheng; Shibo Li; Roy R. Zhang; Wei R. Chen; Hong Liu
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Paper Abstract

The purpose of this study is to investigate the impact of the depth of field (DOF) of microscopic systems on cytogenetic image qualities. Due to the narrow DOF of high magnification, large numerical aperture (N.A.) objective lenses, random vibrations of even high precision scanning stages may result in large amount of off focused images. In this study, the DOF of microscopic systems with various objective magnifications/numerical apertures (N.A.) is first measured using standard resolution targets. The impact of DOF on cytogenetic image qualities is then subjectively evaluated with clinical samples, by comparing the band shape and sharpness of analyzable chromosomes. For a specific digital microscopic system with 100× objective lens (N.A. = 1.25), the results of observational studies revealed that chromosomal bands are still recognizable when the images are obtained approximately +/- 1 μm from the focusing plane. The chromosomal bands become fuzzy and unrecognizable when the system is 1.5 μm away from the focusing position. The results of this preliminary experimental study may provide useful design trade-off parameters for developing optimal scanning microscopic systems for cytogenetic applications.

Paper Details

Date Published: 10 February 2011
PDF: 5 pages
Proc. SPIE 7900, Biophotonics and Immune Responses VI, 79000D (10 February 2011); doi: 10.1117/12.881453
Show Author Affiliations
Yuchen Qiu, Univ. of Oklahoma (United States)
Xiaodong Chen, Tianjin Univ. (China)
Univ. of Oklahoma (United States)
Yuhua Li, Univ. of Oklahoma (United States)
Bin Zheng, Univ. of Pittsburgh (United States)
Shibo Li, Univ. of Oklahoma Health Sciences Ctr. (United States)
Roy R. Zhang, Univ. of Oklahoma Health Sciences Ctr. (United States)
Wei R. Chen, Univ. of Central Oklahoma (United States)
Hong Liu, Univ. of Oklahoma (United States)

Published in SPIE Proceedings Vol. 7900:
Biophotonics and Immune Responses VI
Wei R. Chen, Editor(s)

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