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10 February 2011 The impact of the depth of field on cytogenetic image quality in scanning microscopy
Yuchen Qiu, Xiaodong Chen, Yuhua Li, Bin Zheng, Shibo Li, Roy R. Zhang, Wei R. Chen, Hong Liu
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Proceedings Volume 7900, Biophotonics and Immune Responses VI; 79000D (2011) https://doi.org/10.1117/12.881453
Event: SPIE BiOS, 2011, San Francisco, California, United States
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.
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Yuchen Qiu, Xiaodong Chen, Yuhua Li, Bin Zheng, Shibo Li, Roy R. Zhang, Wei R. Chen, and Hong Liu "The impact of the depth of field on cytogenetic image quality in scanning microscopy", Proc. SPIE 7900, Biophotonics and Immune Responses VI, 79000D (10 February 2011); https://doi.org/10.1117/12.881453
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KEYWORDS
Objectives
Image quality
Lenses
Statistical analysis
Image quality standards
Imaging systems
Microscopy
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