RECONSTRUCTED 3-DIMENSIONAL IMAGES OF FLOW-SORTED NUCLEI HYBRIDIZED WITH CHROMOSOME-SPECIFIC PROBES

We demonstrated that the three-dimensional (3-D) locational and morpho logical differences of chromosome 17 are dependent on each cell cycle phase in the clinical materials, Cell suspensions prepared from hypert rophied tonsil were hybridized with chromosome 17 whole painting probe or its centromeric probe and the probes were detected with fluorescei n isothiocyanate. Then the cells were sorted from G(0 + 1), S-, and G( 2 + M)-phase fractions by flow cytometry and observed by confocal lase r scanning microscopy to obtain the serial optical sections. The 3-D i mages were obtained by assembling these sections using a computerized image analysis device, The distribution of centromeric copies was anal yzed statistically, and the data values were not a population of rando m distribution within a sphere, The copies were observed in the periph ery of the nuclei in G(0 + 1)- and S-phase, The 3-D images revealed th at chromosome 17 was oval in shape in the G(0 + 1)-phase nucleus, and was changing into a flame shape in the S-phase, with arms stretching o ut along the nuclear membrane, and looked bush shaped in G(2)-phase, T he eccentric distribution of chromosome 17 in G(0 + 1)- and S-phase nu clei may reflect the optimal efficiency of incorporating and/or releas ing essential materials and products.