Abstract
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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