Research Papers: Imaging

Impact of the optical depth of field on cytogenetic image quality

[+] Author Affiliations
Yuchen Qiu, Yuhua Li, Hong Liu

University of Oklahoma, Center for Bioengineering and School of Electrical and Computer Engineering, 660 Parrington Oval, Norman, Oklahoma 73019

Xiaodong Chen

University of Oklahoma, Center for Bioengineering and School of Electrical and Computer Engineering, 660 Parrington Oval, Norman, Oklahoma 73019

Tianjin University, College of Precision Instrument and Opto-electronics Engineering, Weijin Road, Tianjin 300072, China

Bin Zheng

University of Pittsburgh, Department of Radiology, 3362 Fifth Avenue, Pittsburgh, Pennsylvania 15213

Shibo Li

University of Oklahoma Health Sciences Center, Department of Pediatrics, 1100 N. Lindsay, Oklahoma City, Oklahoma 73104

Wei R. Chen

University of Central Oklahoma, Department of Engineering and Physics, 100 North University Drive, Edmond, Oklahoma 73034

J. Biomed. Opt. 17(9), 096017 (Sep 18, 2012). doi:10.1117/1.JBO.17.9.096017
History: Received March 23, 2012; Revised August 13, 2012; Accepted August 23, 2012
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Abstract.  In digital pathology, clinical specimen slides are converted into digital images by microscopic image scanners. Since random vibration and mechanical drifting are unavoidable on even high-precision moving stages, the optical depth of field (DOF) of microscopic systems may affect image quality, in particular when using an objective lens with high magnification power. The DOF of a microscopic system was theoretically analyzed and experimentally validated using standard resolution targets under 60× dry and 100× oil objective lenses, respectively. Then cytogenetic samples were imaged at in-focused and off-focused states to analyze the impact of DOF on the acquired image qualities. For the investigated system equipped with the 60× dry and 100× oil objective lenses, the theoretical estimation of the DOF are 0.855 μm and 0.703 μm, and the measured DOF are 3.0 μm and 1.8 μm, respectively. The observation reveals that the chromosomal bands of metaphase cells are distinguishable when images are acquired up to approximately 1.5 μm or 1 μm out of focus using the 60× dry and 100× oil objective lenses, respectively. The results of this investigation provide important designing trade-off parameters to optimize the digital microscopic image scanning systems in the future.

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© 2012 Society of Photo-Optical Instrumentation Engineers

Citation

Yuchen Qiu ; Xiaodong Chen ; Yuhua Li ; Bin Zheng ; Shibo Li, et al.
"Impact of the optical depth of field on cytogenetic image quality", J. Biomed. Opt. 17(9), 096017 (Sep 18, 2012). ; http://dx.doi.org/10.1117/1.JBO.17.9.096017


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