M. Matsuta et al., RECONSTRUCTED 3-DIMENSIONAL IMAGES OF FLOW-SORTED NUCLEI HYBRIDIZED WITH CHROMOSOME-SPECIFIC PROBES, The Journal of histochemistry and cytochemistry, 44(11), 1996, pp. 1337-1343
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.