Human archival tissues provide a valuable source for the analysis of spatial genome organization.
Sections from archival formalin-fixed, paraffin wax-embedded human tissues are a valuable source for the study of the nuclear architecture of specific tissue types in terms of the three-dimensional spatial positioning and architecture of chromosome territories and sub-chromosomal domains. Chromosome painting, centromeric, and locus-specific probes were hybridized to tissue microarrays prepared from formalin-fixed paraffin wax-embedded samples of pancreas and breast. The cell nuclei were analyzed using quantitative three-dimensional image microscopy. The results obtained from non-neoplastic pancreatic cells of randomly selected individuals indicated that the radial arrangement of the chromosome 8 territories as well as their shape (roundness) did not significantly differ between the individuals and were in accordance with assumptions of a probabilistic model for computer simulations. There were considerable differences between pancreatic tumor and non-neoplastic cells. In non-neoplastic ductal epithelium of the breast there was a larger, but insignificant, variability in the three-dimensional positioning of the centromere 17 and HER2 domains between individuals. In neoplastic epithelial breast cells, however, the distances between centromere and gene domains were, on average, smaller than in non-neoplastic cells. In conclusion, our results demonstrate the feasibility of studying the genome architecture in archival, formalin-fixed, paraffin wax-embedded human tissues, opening new directions in tumor research and cell classification.
Wiech
Thorsten
T
Institute of Pathology, University Hospital Freiburg, Albertstrasse 19, 79104 Freiburg, Germany.
Timme
Sylvia
S
Riede
Florian
F
Stein
Stefan
S
Schuricke
Michael
M
Cremer
Christoph
C
Werner
Martin
M
Hausmann
Michael
M
Walch
Axel
A
D001940:Breast / Q000033:anatomy & histology; D001943:Breast Neoplasms / Q000235:genetics / Q000473:pathology*; D018270:Carcinoma, Ductal, Breast / Q000235:genetics / Q000473:pathology*; D021441:Carcinoma, Pancreatic Ductal / Q000235:genetics / Q000473:pathology*; D045584:Chromosome Positioning; D004848:Epithelium / Q000033:anatomy & histology; D015894:Genome, Human; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D010183:Pancreatic Ducts / Q000033:anatomy & histology; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000473:pathology*; D016707:Tissue Fixation