Automatic segmentation of cell nuclei in Feulgen-stained histological sections of prostate cancer and quantitative evaluation of segmentation results

Cytometry A. 2012 Jul;81(7):588-601. doi: 10.1002/cyto.a.22068. Epub 2012 May 17.


Digital image analysis of cell nuclei is useful to obtain quantitative information for the diagnosis and prognosis of cancer. However, the lack of a reliable automatic nuclear segmentation is a limiting factor for high-throughput nuclear image analysis. We have developed a method for automatic segmentation of nuclei in Feulgen-stained histological sections of prostate cancer. A local adaptive thresholding with an object perimeter gradient verification step detected the nuclei and was combined with an active contour model that featured an optimized initialization and worked within a restricted region to improve convergence of the segmentation of each nucleus. The method was tested on 30 randomly selected image frames from three cases, comparing the results from the automatic algorithm to a manual delineation of 924 nuclei. The automatic method segmented a few more nuclei compared to the manual method, and about 73% of the manually segmented nuclei were also segmented by the automatic method. For each nucleus segmented both manually and automatically, the accuracy (i.e., agreement with manual delineation) was estimated. The mean segmentation sensitivity/specificity were 95%/96%. The results from the automatic method were not significantly different from the ground truth provided by manual segmentation. This opens the possibility for large-scale nuclear analysis based on automatic segmentation of nuclei in Feulgen-stained histological sections.

MeSH terms

  • Algorithms
  • Automation, Laboratory
  • Cell Nucleus / pathology*
  • Cell Nucleus / physiology
  • Cell Nucleus Size
  • Coloring Agents / chemistry*
  • Humans
  • Image Interpretation, Computer-Assisted*
  • Male
  • Prostatic Neoplasms / diagnosis
  • Prostatic Neoplasms / pathology*
  • ROC Curve
  • Rosaniline Dyes / chemistry*
  • Staining and Labeling


  • Coloring Agents
  • Feulgen stain
  • Rosaniline Dyes