Automated segmentation and analysis of fluorescent in situ hybridization (FISH) signals in interphase nuclei of pap-smear specimens

Xingwei Wang; Bin Zheng; Shibo Li; Roy R. Zhang; Yuhua Li; John J. Mulvihill; Wei R. Chen; Hong Liu

doi:10.1117/12.813709

12 February 2009 Automated segmentation and analysis of fluorescent in situ hybridization (FISH) signals in interphase nuclei of pap-smear specimens

Xingwei Wang, Bin Zheng, Shibo Li, Roy R. Zhang, Yuhua Li, John J. Mulvihill, Wei R. Chen, Hong Liu

Author Affiliations +

Proceedings Volume 7176, Dynamics and Fluctuations in Biomedical Photonics VI; 717609 (2009) https://doi.org/10.1117/12.813709
Event: SPIE BiOS, 2009, San Jose, California, United States

Abstract

Interphase fluorescence in situ hybridization (FISH) technology is a potential and promising molecular imaging tool, which can be applied to screen and detect cervical cancer. However, manual FISH detection method is a subjective, tedious, and time-consuming process that results in a large inter-reader variability and possible detection error (in particular for heterogeneous cases). Automatic FISH image analysis aims to potentially improve detection efficiency and also produce more accurate and consistent results. In this preliminary study, a new computerized scheme is developed to automatically segment analyzable interaphase cells and detect FISH signals using digital fluorescence microscopic images acquired from Pap-smear specimens. First, due to the large intensity variations of the acquired interphase cells and overlapping cells, an iterative (multiple) threshold method and a feature-based classifier are applied to detect and segment all potentially analyzable interphase nuclei depicted on a single image frame. Second, a region labeling algorithm followed up a knowledge-based classifier is implemented to identify splitting and diffused FISH signals. Finally, each detected analyzable cell is classified as normal or abnormal based on the automatically counted number of FISH signals. To test the performance of this scheme, an image dataset involving 250 Pap-smear FISH image frames was collected and used in this study. The overall accuracy rate for segmenting analyzable interphase nuclei is 86.6% (360/424). The sensitivity and specificity for classifying abnormal and normal cells are 88.5% and 86.6%, respectively. The overall cell classification agreement rate between our scheme and a cytogeneticist is 86.6%. The testing results demonstrate the feasibility of applying this automated scheme in FISH image analysis.

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Xingwei Wang, Bin Zheng, Shibo Li, Roy R. Zhang, Yuhua Li, John J. Mulvihill, Wei R. Chen, and Hong Liu "Automated segmentation and analysis of fluorescent in situ hybridization (FISH) signals in interphase nuclei of pap-smear specimens", Proc. SPIE 7176, Dynamics and Fluctuations in Biomedical Photonics VI, 717609 (12 February 2009); https://doi.org/10.1117/12.813709

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