Three‐dimensional distribution of centromeric or paracentromeric heterochromatin of chromosomes 1, 7, 15 and 17 in human lymphocyte nuclei studied with light microscopic axial tomography

Light microscopic axial tomography was applied to examine the position of peri- or paracentromeric chromosomal targets in nuclei of PHA-stimulated human lymphocytes following two color fluorescence in situ hybridization with DNA probes for 1q12 and 15p or 7c and 17c. Evaluation of hybridized nuclei was performed in a glass capillary device. By turning the capillary around its longitudinal axis each nucleus could be viewed at any desired angle. To measure the true three-dimensional distance between two chromosomal targets, the nucleus was turned around until both targets were positioned in the same focal plane. Similarly, the true distance between a target region and the center of the nucleus was estimated. The results provide evidence for differences in the three-dimensional nuclear distribution of the target regions. In particular, 7c was positioned more peripherally than the other chromosomal targets. Experimental data were compared with several models for a distribution of chromosomal targets under topological constraints. These models take into account spatial limitations of the distribution of the chromosome territories which harbor the hybridized targets.

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