Abstract
The analysis of cell characteristics from high-resolution digital histopathological images is the standard clinical practice for the diagnosis and prognosis of cancer. Yet, it is a rather exhausting process for pathologists to examine the cellular structures manually in this way. Automating this tedious and time-consuming process is an emerging topic of the histopathological image-processing studies in the literature. This paper presents a two-stage segmentation method to obtain cellular structures in high-dimensional histopathological images of renal cell carcinoma. First, the image is segmented to superpixels with simple linear iterative clustering (SLIC) method. Then, the obtained superpixels are clustered by the state-of-the-art clustering-based segmentation algorithms to find similar superpixels that compose the cell nuclei. Furthermore, the comparison of the global clustering-based segmentation methods and local region-based superpixel segmentation algorithms are also compared. The results show that the use of the superpixel segmentation algorithm as a pre-segmentation method improves the performance of the cell segmentation as compared to the simple single clustering-based segmentation algorithm. The true positive ratio (TPR), true negative ratio (TNR), F-measure, precision, and overlap ratio (OR) measures are utilized as segmentation performance evaluation. The computation times of the algorithms are also evaluated and presented in the study.

The visual flowchart of the proposed automatic cell segmentation in histopathological images via two-staged superpixel-based algorithms.








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Achanta R, Shaji A, Smith K, Lucchi A, Fua P, Süsstrunk S (2012) Slic superpixels compared to state-of-the-art superpixel methods. IEEE Trans Pattern Anal Mach Intell 34(11):2274–2282
Akbar S, Jordan L, Thompson AM, McKenna SJ (2015) Tumor localization in tissue microarrays using rotation invariant superpixel pyramids. In: IEEE 12th International Symposium on Biomedical Imaging, ISBI’15, IEEE, pp 1292–1295
Al-Lahham H, Alomari R, Hiary H, Chaudhary V (2012) Automation proliferation rate estimation from breast cancer ki-67 histology images. Proceedings of the SPIE Medical Imaging: Computer-Aided Diagnosis 8315 83:152A
Ali S, Lewis J, Madabhushi A (2013) Spatially aware cell cluster (SPACCL) graphs: Predicting outcome in oropharyngeal p16+ tumors. In: International Conference on Medical Image Computing and Computer-Assisted Intervention, MICCAI’13, Springer, pp 412–419
Beck AH, Sangoi AR, Leung S, Marinelli RJ, Nielsen TO, van de Vijver MJ, West RB, van de Rijn M, Koller D (2011) Systematic analysis of breast cancer morphology uncovers stromal features associated with survival. Sci Transl Med 3(108):108ra113–108ra113
Van den Bergh M, Van Gool L (2012) Real-time stereo and flow-based video segmentation with superpixels. In: IEEE Workshop on Applications of Computer Vision, WACV’12, IEEE, pp 89–96
Bezdek JC (2013) Pattern recognition with fuzzy objective function algorithms. Springer Science & Business Media, Berlin
Cheng J, Liu J, Xu Y, Yin F, Wong DWK, Tan NM, Tao D, Cheng CY, Aung T, Wong TY (2013) Superpixel classification based optic disc and optic cup segmentation for glaucoma screening. IEEE Trans Med Imaging 32(6):1019–1032
Cheng X, Wang Y, Yuan X, Li B, Ding Y, Zhang Z (2015) Improving video foreground segmentation and propagation through multifeature fusion. J Electron Imaging 24(6):063,017–063,017
Du M, Wu X, Chen W, Wang J (2016) Exploiting multiple contexts for saliency detection. J Electron Imaging 25(6):063,005–063,005
Dunn JC (1973) A fuzzy relative of the isodata process and its use in detecting compact well-separated clusters. J Cybern 3:32–57
Ester M, Kriegel HP, Sander J, Xu X et al (1996) A density-based algorithm for discovering clusters in large spatial databases with noise. In: KDD, vol 96-34, pp 226–231
George YM, Bagoury BM, Zayed HH, Roushdy MI (2013) Automated cell nuclei segmentation for breast fine needle aspiration cytology. Signal Process 93(10):2804–2816
Irshad H, Montaser-Kouhsari L, Waltz G, Bucur O, Nowak J, Dong F, Knoblauch NW, Beck AH (2014) Crowdsourcing image annotation for nucleus detection and segmentation in computational pathology: evaluating experts, automated methods, and the crowd. In: Pacific Symposium on Biocomputing, PSB’15, NIH Public Access, pp 294–305
Kovesi P (2013) Image segmentation using SLIC superpixels and DBSCAN clustering. http://www.peterkovesi.com/projects/segmentation/index.html, accessed: 2017-04-22
Kovesi PD (2000) Matlab and octave functions for computer vision and image processing. Online: http://www.csseuwaeduau/∼pk/Research/MatlabFns/#match
Liu F, Lin G, Shen C (2015) CRF learning with CNN features for image segmentation. Pattern Recognit 48(10):2983–2992
Liu MY, Tuzel O, Ramalingam S, Chellappa R (2011) Entropy rate superpixel segmentation. In: IEEE Conference on Computer Vision and Pattern Recognition, CVPR’11, IEEE, pp 2097–2104
Lu C, Mahmood M, Jha N, Mandal M (2012) A robust automatic nuclei segmentation technique for quantitative histopathological image analysis. Anal Quant Cytol Histol 34:296–308
MacQueen J et al (1967) Some methods for classification and analysis of multivariate observations. In: Proceedings of the 5th Berkeley Symposium on Mathematical Statistics and Probability, Oakland, CA, USA., vol 1-14, pp 281-297
Malamateniou C, Rutherford M, Hajnal JV, Glocker B, Rueckert D (2015) Automatic brain localization in fetal MRI using superpixel graphs. In: Machine learning meets medical imaging: 1st international workshop, MLMMI’15, conjunction with ICML 2015, Lille, France, July 11, 2015, revised selected papers, Springer, vol 9487, p 13
Meng F, Li H, Liu G, Ngan KN (2012) Object co-segmentation based on shortest path algorithm and saliency model. IEEE Trans Multimedia 14(5):1429–1441
Ochs P, Malik J, Brox T (2014) Segmentation of moving objects by long-term video analysis. IEEE Trans Pattern Anal Mach Intell 36(6):1187–1200
Schick A, Bäuml M, Stiefelhagen R (2012) Improving foreground segmentations with probabilistic superpixel Markov random fields. In: IEEE Computer Society Conference on Computer Vision and Pattern Recognition Workshops, CVPRW’12, IEEE, pp 27–31
Shen P, Qin W, Yang J, Hu W, Chen S, Li L, Wen T, Gu J (2015) Segmenting multiple overlapping nuclei in H&E stained breast cancer histopathology images based on an improved watershed. In: 2015 IET Int. Conference on Biomedical Image and Signal Processing, ICBISP’15, IET, pp 1–4
Sirinukunwattana K, Snead DR, Rajpoot NM (2015) A novel texture descriptor for detection of glandular structures in colon histology images. In: SPIE Med Imaging, International Society for Optics and Photonics, pp 94,200S–94,200S
Sun F, Qin K, Sun W, Guo H (2016) Fast background subtraction for moving cameras based on nonparametric models. J Electron Imaging 25(3):033,017–033,017
Tang D, Fu H, Cao X (2012) Topology preserved regular superpixel. In: IEEE International Conference on Multimedia and Expo ICME’12, IEEE, pp 765–768
Wright AI, Magee D, Quirke P, Treanor D (2016) Incorporating local and global context for better automated analysis of colorectal cancer on digital pathology slides. Procedia Comput Sci 90:125–131
Xing F, Yang L (2013) Robust cell segmentation for non-small cell lung cancer. In: IEEE 10th International Symposium on Biomedical Imaging, ISBI’13, IEEE, pp 386–389
Xu H, Lu C, Mandal M (2014) An efficient technique for nuclei segmentation based on ellipse descriptor analysis and improved seed detection algorithm. IEEE J Biomed Health Inf 18(5):1729–1741
Xu J, Xiang L, Liu Q, Gilmore H, Wu J, Tang J, Madabhushi A (2016) Stacked sparse autoencoder (SSAE) for nuclei detection on breast cancer histopathology images. IEEE Trans Med Imaging 35(1):119–130
Yang F, Lu H, Yang MH (2014) Robust superpixel tracking. IEEE Trans Image Process 23(4):1639–1651
Acknowledgements
This work was supported by the Scientific Research Projects Coordination Department, Yildiz Technical University, under Project 2014-04-01-KAP01. The authors also would like to thank Beck Laboratory at Harvard University for providing and annotating the high-resolution histopathological images of renal cell carcinoma data set selected from the Cancer Genome Atlas (TCGA) data portal and publicly available for academic usage. The authors state no conflicts of interest and have nothing to disclose.
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Albayrak, A., Bilgin, G. Automatic cell segmentation in histopathological images via two-staged superpixel-based algorithms. Med Biol Eng Comput 57, 653–665 (2019). https://doi.org/10.1007/s11517-018-1906-0
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DOI: https://doi.org/10.1007/s11517-018-1906-0