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

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.

Fluorescent dot counting in interphase cell nuclei

“…There is evidence that chromosomes occupy distinct territories in cell nuclei. Dietzel et al have shown a significant difference between observed data and this model (Dietzel et al 1995). But the differences are small, especially when the distance between the targets is small.…”

Fluorescent dot counting in interphase cell nuclei

“…There is evidence that chromosomes occupy distinct territories in cell nuclei. Dietzel et al have shown a significant difference between observed data and this model (Dietzel et al 1995). But the differences are small, especially when the distance between the targets is small.…”

Efficient, interactive, and three-dimensional segmentation of cell nuclei in thick tissue sections

“…In conjunction with stoichiometric DNA staining and quantitative image acquisition, our method could be used for measuring the DNA content of individual nuclei in tissue sections (7,12,29). Similarly, in conjunction with techniques for automatically (27) or interactively (17) detecting punctate FISH signals, our method could be used for enumerating FISH signals per nucleus and for measuring the spatial positions of them within each nucleus (8,11,14).…”

The spatial distribution of human immunoglobulin genes within the nucleus: evidence for gene topography independent of cell type and transcriptional activity

“…Possibly, the variability observed in the distributions of all genes analyzed may reflect either variations in the positioning of the respective chromosome or actual gene movements (or both), within a constrained nuclear subvolume (see Dietzel et al 1995;Münkel et al 1995). Therefore, an important and still unanswered question will be whether gene activity can influence subtle gene movements within a defined topographical region.…”

Fluorescent dot counting in interphase cell nuclei

“…There is evidence that chromosomes occupy distinct territories in cell nuclei. Dietzel et al have shown a significant difference between observed data and this model (Dietzel et al 1995). But the differences are small, especially when the distance between the targets is small.…”

Efficient, interactive, and three-dimensional segmentation of cell nuclei in thick tissue sections

“…In conjunction with stoichiometric DNA staining and quantitative image acquisition, our method could be used for measuring the DNA content of individual nuclei in tissue sections (7,12,29). Similarly, in conjunction with techniques for automatically (27) or interactively (17) detecting punctate FISH signals, our method could be used for enumerating FISH signals per nucleus and for measuring the spatial positions of them within each nucleus (8,11,14).…”

The spatial distribution of human immunoglobulin genes within the nucleus: evidence for gene topography independent of cell type and transcriptional activity

“…Possibly, the variability observed in the distributions of all genes analyzed may reflect either variations in the positioning of the respective chromosome or actual gene movements (or both), within a constrained nuclear subvolume (see Dietzel et al 1995;Münkel et al 1995). Therefore, an important and still unanswered question will be whether gene activity can influence subtle gene movements within a defined topographical region.…”

Fluorescent dot counting in interphase cell nuclei

“…There is evidence that chromosomes occupy distinct territories in cell nuclei. Dietzel et al have shown a significant difference between observed data and this model (Dietzel et al 1995). But the differences are small, especially when the distance between the targets is small.…”

Efficient, interactive, and three-dimensional segmentation of cell nuclei in thick tissue sections

“…In conjunction with stoichiometric DNA staining and quantitative image acquisition, our method could be used for measuring the DNA content of individual nuclei in tissue sections (7,12,29). Similarly, in conjunction with techniques for automatically (27) or interactively (17) detecting punctate FISH signals, our method could be used for enumerating FISH signals per nucleus and for measuring the spatial positions of them within each nucleus (8,11,14).…”

The spatial distribution of human immunoglobulin genes within the nucleus: evidence for gene topography independent of cell type and transcriptional activity

“…Possibly, the variability observed in the distributions of all genes analyzed may reflect either variations in the positioning of the respective chromosome or actual gene movements (or both), within a constrained nuclear subvolume (see Dietzel et al 1995;Münkel et al 1995). Therefore, an important and still unanswered question will be whether gene activity can influence subtle gene movements within a defined topographical region.…”