COMBO-FISH for focussed fluorescence labelling of gene domains: 3D-analysis of the genome architecture of abl and bcr in human blood cells

Jutta Schwarz-Finsterle, Stefan Stein, Constance Großmann, Eberhard Schmitt, Helmut Schneider, Luba Trakhtenbrot, Gideon Rechavi, Ninette Amariglio, Christoph Cremer, Michael Hausmann

Research output: Contribution to journalArticlepeer-review

Abstract

Structural analysis and nanosizing of gene domains requires not only high-resolution microscopy but also improved techniques of fluorescence labelling strongly focussed on the gene domains. To investigate the architecture of abl and bcr in blood cell nuclei forming the Philadelphia chromosome in CML, we applied COMBO-FISH using specifically colocalising combinations of triple strand forming oligonucleotide probes for abl on chromosome 9 and bcr on chromosome 22. Each probe set consisting of 31 homopyrimidine oligonucleotides was computer selected from the human genome database. Measurements by 3D microscopy were compared to results obtained after standard FISH using commercially available abl/bcr BAC probes. The relative radial fluorescence distributions in lymphocyte cell nuclei of healthy donors in comparison to cell nuclei of blood cells of CML patients showed a strong correlation in the location of abl and bcr for both labelling techniques. The absolute distances of the homologous bcr domains and the abl domain - nuclear center - abl domain angles in cell nuclei of CML donors differed significantly from those of healthy donors only when COMBO-FISH was applied. These results indicate that COMBO-FISH may be more sensitive than standard FISH in case of slight modifications in the genome architecture.

Original languageEnglish
Pages (from-to)1038-1046
Number of pages9
JournalCell Biology International
Volume29
Issue number12
DOIs
StatePublished - Dec 2005
Externally publishedYes

Keywords

  • 3D confocal laser scanning microscopy
  • COMBO-FISH
  • FISH
  • Genome architecture
  • Image analysis

Fingerprint

Dive into the research topics of 'COMBO-FISH for focussed fluorescence labelling of gene domains: 3D-analysis of the genome architecture of abl and bcr in human blood cells'. Together they form a unique fingerprint.

Cite this