Telomeres play a critical role in the maintenance of chromosomal stability. Telomere erosion, coupled with loss of DNA
damage checkpoint function, results in genomic instability that promotes the development of cancer. The critical role of
telomere dynamics in cancer has motivated the development of technologies designed to monitor telomere reserves in a
highly quantitative and high-throughput manner in humans and model organisms. To this end, we have adapted and
modified two established technologies, telomere-FISH and laser scanning cytometry. Specifically, we have produced a
number of enhancements to the iCys LSC (CompuCyte) package including software updates, use of 60X dry objectives,
and increased spatial resolution by 0.2 um size of stage steps. In addition, the 633 nm HeNe laser was replaced with a
532 nm green diode laser to better match the viewing options. Utilization of telomere-deficient mouse cells with short
dysfunctional telomeres and matched telomerase reconstituted cultures demonstrated significantly higher mean integral
specific fluorescence values for mTR transfectants relative to empty vector controls: 4.485M vs. 1.362M (p<0.0001).
Histograms of average telomere intensities for individual cells were obtained and demonstrated intercellular
heterogeneity in telomere lengths. The validation of the approach derives from a strong correlation between iCys LSC
values and Southern blotting. This validated method greatly increases our experimental throughput and objectivity.