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Biophys J, November 1999, p. 2871-2886, Vol. 77, No. 5
*Institute of Applied Physics,
The motion of subchromosomal foci and of whole chromosome
territories in live human cell nuclei was investigated in
four-dimensional space-time images. Visualization of subchromosomal
foci was achieved by incorporating Cy3-dUTP into the nuclear DNA of two
different cell types after microinjection. A subsequent segregation of
the labeled cell nuclei led to the presence of only a few labeled chromosome territories on a background of nonlabeled chromatin (Zink et
al., 1998. Hum. Genet. 102:241-251). This procedure
yielded many distinct signals in a given cell nucleus. Motion analysis in four-dimensional space-time images was performed using
single-particle tracking and a statistical approach to the detection of
a possible directional motion of foci relative to the center of mass of
a chromosome territory. The accuracy of the analysis was tested using
simulated data sets that closely mirrored the experimental setup and
using microparticles of known size. Application of the analysis tools
to experimental data showed that mutual diffusion-like movements
between foci located on different chromosomes were more pronounced than
inside the territories. In the time range observed, movements of
individual foci could best be described by a random diffusion process.
The statistical test for joint directed motion of several foci inside
chromosome territories revealed that foci occasionally switched from
random to directional motion inside the territories.
Biophys J, November 1999, p. 2871-2886, Vol. 77, No. 5
© 1999 by the Biophysical Society 0006-3495/99/11/2871/16 $2.00
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