Open AccessThree‐dimensional reconstruction of pericentromeric (1q12) DNA and ribosomal RNA sequences in HL60 cells after double‐target in situ hybridization and confocal microscopy
Author(s) -
van Dekken H.,
van der Voort H. T. M.,
Brakenhoff G. J.,
Bauman J. G. J.
Publication year - 1990
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/cyto.990110504
Subject(s) - ribosomal rna , biology , in situ hybridization , in situ , ribosomal dna , microbiology and biotechnology , dna , hybridization probe , confocal , fluorescence in situ hybridization , cytoplasm , centromere , rna , molecular probe , confocal microscopy , chromosome , gene , gene expression , genetics , chemistry , optics , phylogenetics , physics , organic chemistry
A fluorescent in situ hybridization procedure was applied to simultaneously label intranuclear pericentromeric (1q12) sequences of the chromosomes 1 and cytoplasmic ribosomal RNA sequences in whole cells of the promyelocytic HL60 cell line. For this purpose biotinated chromosome 1‐specific (1q12) repetitive satellite DNA and 28S ribosomal ssRNA probes were used. The entire procedure was performed in suspension to preserve nuclear morphology. The result was studied by three‐dimensional analysis, as provided by a scanning laser confocal microscope. The intracellular positions of both cytoplasmic rRNA and intranuclear centromere 1 DNA could easily be distinguished. This approach could be useful as a framework for the study of the 3‐D localization of genes and gene transcripts.