Translocation of genes into the pericentromeric heterochromatin occurs during cellular differentiation and leads to a long-term silencing of these genes. Consequently, a structural remodelling of this heterochromatin compartment is observed during differentiation but this remains to be defined from a topological point of view. In a previous study, we analysed the three-dimensional (3D) distribution patterns of centromere clusters (chromocentres) by confocal scanning laser microscopy and found that differentiation of the promyelocytic leukaemia cell line NB4 along the neutrophil lineage is associated with a progressive clustering of centromeres. This clustering was reflected by a decreased number of detectable chromocentres, i.e. groups of centromeres with a distance below the diffraction-limited resolution of optical microscopy. The purpose of this study was to perform a statistical analysis of the 3D distribution of chromocentres in NB4 cells. Several point field characteristics (Ripley's K-function, L-function, pair correlation function, nearest-neighbour distribution function) were investigated to describe the topology of chromocentres during differentiation of NB4 cells. The pair correlation function revealed a higher frequency of chromocentre distances between 350 nm and 800 nm in undifferentiated NB4 cells as compared with differentiated cells. The L-function and the nearest-neighbour distribution function confirmed these results. These data imply the existence of intranuclear heterochromatin zones formed by functionally related centromeric regions. In view of the observed decrease in the number of detectable chromocentres during differentiation, we hypothesize that these zones with a diameter of 350-800 nm in undifferentiated NB4 cells contract into zones with a diameter below 350 nm in differentiated cells.