Graph cut and image intensity-based splitting improves nuclei segmentation in high-content screening

Quantification of phenotypes in high-content screening experiments depends on the accuracy of single cell analysis. In such analysis workflows, cell nuclei segmentation is typically the first step and is followed by cell body segmentation, feature extraction, and subsequent data analysis workflows. Therefore, it is of utmost importance that the first steps of high-content analysis are done accurately in order to guarantee correctness of the final analysis results. In this paper, we present a novel cell nuclei image segmentation framework which exploits robustness of graph cut to obtain initial segmentation for image intensity-based clump splitting method to deliver the accurate overall segmentation. By using quantitative benchmarks and qualitative comparison with real images from high-content screening experiments with complicated multinucleate cells, we show that our method outperforms other state-of-the-art nuclei segmentation methods. Moreover, we provide a modular and easy-to-use implementation of the method for a widely used platform.

[1]  Anne E Carpenter,et al.  A call for bioimaging software usability , 2012, Nature Methods.

[2]  D. Greig,et al.  Exact Maximum A Posteriori Estimation for Binary Images , 1989 .

[3]  Anne E Carpenter,et al.  CellProfiler: image analysis software for identifying and quantifying cell phenotypes , 2006, Genome Biology.

[4]  Gareth Funka-Lea,et al.  Graph Cuts and Efficient N-D Image Segmentation , 2006, International Journal of Computer Vision.

[5]  Carlos Ortiz-de-Solorzano,et al.  A Two-Phase Segmentation of Cell Nuclei Using Fast Level Set-Like Algorithms , 2009, SCIA.

[6]  Vladimir Kolmogorov,et al.  An experimental comparison of min-cut/max- flow algorithms for energy minimization in vision , 2001, IEEE Transactions on Pattern Analysis and Machine Intelligence.

[7]  P. Liberali,et al.  Population context determines cell-to-cell variability in endocytosis and virus infection , 2009, Nature.

[8]  Pauli Rämö,et al.  CellClassifier: supervised learning of cellular phenotypes , 2009, Bioinform..

[9]  Pekka Ruusuvuori,et al.  Open Access Research Article Evaluation of Methods for Detection of Fluorescence Labeled Subcellular Objects in Microscope Images , 2022 .

[10]  C Chen,et al.  Constraint factor graph cut–based active contour method for automated cellular image segmentation in RNAi screening , 2008, Journal of microscopy.

[11]  Carlos Ortiz-de-Solorzano,et al.  Segmentation of Touching Cell Nuclei Using a Two-Stage Graph Cut Model , 2009, SCIA.

[12]  Carolina Wählby,et al.  Image Based Measurements of Single Cell mtDNA Mutation Load , 2007, SCIA.

[13]  Wiro J. Niessen,et al.  Advanced level-set based multiple-cell segmentation and tracking in time-lapse fluorescence microscopy images , 2008, 2008 5th IEEE International Symposium on Biomedical Imaging: From Nano to Macro.

[14]  K. Rohr,et al.  Single‐cell‐based image analysis of high‐throughput cell array screens for quantification of viral infection , 2009, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[15]  Vladimir Kolmogorov,et al.  Computing geodesics and minimal surfaces via graph cuts , 2003, Proceedings Ninth IEEE International Conference on Computer Vision.

[16]  Joakim Lindblad,et al.  Image analysis for automatic segmentation of cytoplasms and classification of Rac1 activation , 2004, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[17]  C Wählby,et al.  Combining intensity, edge and shape information for 2D and 3D segmentation of cell nuclei in tissue sections , 2004, Journal of microscopy.

[18]  Chunming Li,et al.  Distance Regularized Level Set Evolution and Its Application to Image Segmentation , 2010, IEEE Transactions on Image Processing.

[19]  Xiaobo Zhou,et al.  Automatic Segmentation of High-Throughput RNAi Fluorescent Cellular Images , 2008, IEEE Transactions on Information Technology in Biomedicine.

[20]  Olli Yli-Harja,et al.  A novel method for splitting clumps of convex objects incorporating image intensity and using rectangular window-based concavity point-pair search , 2013, Pattern Recognit..