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Measurement of telomere organization using widefield microscopy Duncan, Tracy

Abstract

Widefield microscopy was used to study the location of telomeres in human interphase lymphocyte nuclei in order to determine whether telomeres are required for nuclear organization. To do this, a microscope's computer control system was programmed to automatically focus the microscope, and large numbers of images were analyzed. Cells in cell cycle stages G1 or S/G2 with long, medium, and short telomeres were photographed. Their two-dimensional images were analyzed to calculate the telomere distribution along the radius of the nucleus and to evaluate the degree of telomere clustering in space using Delaunay graphs. Telomeres in nuclei with short telomeres were shown to be distributed closer to the center of the nucleus than those in nuclei with long telomeres. The radial distribution of telomeres did not change with cell cycle phase. No changes in the degree of telomere clustering with telomere length or cell cycle phase were observed.

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