Cell and Tissue Research

, Volume 322, Issue 2, pp 257–267 | Cite as

Nuclear patterns of human breast cancer cells during apoptosis: characterisation by fractal dimension and co-occurrence matrix statistics

Regular Article


An analytical strategy combining fractal geometry and grey-level co-occurrence matrix (GLCM) statistics was devised to investigate ultrastructural changes in oestrogen-insensitive SK-BR3 human breast cancer cells undergoing apoptosis in vitro. Apoptosis was induced by 1 μM calcimycin (A23187 Ca2+ ionophore) and assessed by measuring conventional cellular parameters during the culture period. SK-BR3 cells entered the early stage of apoptosis within 24 h of treatment with calcimycin, which induced detectable changes in nuclear components, as documented by increased values of most GLCM parameters and by the general reduction of the fractal dimensions. In these affected cells, morphonuclear traits were accompanied by the reduction of distinct gangliosides and loss of unidentifiable glycolipid molecules at the cell surface. All these changes were shown to be involved in apoptosis before the detection of conventional markers, which were only measurable during the active phases of apoptotic cell death. In overtly apoptotic cells treated with 1 μM calcimycin for 72 h, most nuclear components underwent dramatic ultrastructural changes, including marginalisation and condensation of chromatin, as reflected in a significant reduction of their fractal dimensions. Hence, both fractal and GLCM analyses confirm that the morphological reorganisation of nuclei, attributable to a loss of structural complexity, occurs early in apoptosis.


Early/late apoptosis Nuclear eu-/hetero-chromatin Fractal dimension Grey-level co-occurrence matrix (GLCM) analysis Surface gangliosides Human breast cancer cells 



fractal dimension


grey-level co-occurrence matrix


fluorescein isothiocyanate


propidium iodide


total lipid extract


perinuclear membrane


nuclear membrane-bound heterochromatin space and outline


scattered heterochromatin space and outline


total heterochromatin space and outline


inter(eu)chromatin space and outline


Terminal deoxynucleotidyl Transferase



Helpful discussions with Prof. Theo Nonnenmacher, University of Ulm, Germany and Prof. Carlo Pellicciari, University of Pavia, Italy are gratefully acknowledged.


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Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  1. 1.Institute for Scientific Interdisciplinary StudiesLocarnoSwitzerland
  2. 2.Faculty of Biology and MedicineUniversity of LausanneLausanneSwitzerland
  3. 3.OCL HospitalLuganoSwitzerland

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