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(American Journal of Pathology. 2003;163:1751-1756.)
© 2003 American Society for Investigative Pathology

Technical Advance

Quantifying Telomere Lengths of Human Individual Chromosome Arms by Centromere-Calibrated Fluorescence in Situ Hybridization and Digital Imaging

Sven Perner*, Silke Brüderlein*, Cornelia Hasel*, Irena Waibel*, Alexandra Holdenried*, Neslisah Ciloglu*, Heiko Chopurian{dagger}, Kirsten Vang Nielsen{ddagger}, Andreas Plesch§, Josef Högel and Peter Möller*

From the Institute of Pathology*and the Women’s Hospital,{dagger}University Hospitals of Ulm, Ulm, Germany; DakoCytomation A/S,{ddagger}Glostrup, Denmark; MetaSystems GmbH,§Altlussheim, Germany; and the Department of Biometrics and Medical Documentation,University of Ulm, Ulm, Germany

Telomere length analysis has aroused considerable interest in biology and oncology. However, most published data are pan-genomic Southern-blot-based estimates. We developed T/C-FISH (telomere/centromere-FISH), allowing precise measurement of individual telomeres at every single chromosome arm. Metaphase preparations are co-hybridized with peptide nucleic acid probes for telomeric sequences and the chromosome 2 centromere serving as internal reference. Metaphase images are captured and karyotyped using dedicated software. A software module determines the absolute integrated fluorescence intensities of the p- and q-telomeres of each chromosome and the reference signal. Normalized data are derived by calculating the ratio of absolute telomere and reference signal intensities, and descriptive statistics are calculated. T/C-FISH detects even small differences in telomere length. Using T/C-FISH we have discovered an epigenetic process occurring in the human male postzygote or early embryo: in umbilical cord blood lymphocytes, telomeres on male Xqs are around 1100 bp shorter than female Xqs.




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