Quantifying telomere lengths of human individual chromosome arms by centromere-calibrated fluorescence in situ hybridization and digital imaging.
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.
Institute of Pathology, University Hospitals of Ulm, Albert-Einstein-Allee 11, D-89081 Ulm, Germany.
Sven
S
Perner
Silke
S
Brüderlein
Cornelia
C
Hasel
Irena
I
Waibel
Alexandra
A
Holdenried
Neslisah
N
Ciloglu
Heiko
H
Chopurian
Kirsten Vang
KV
Nielsen
Andreas
A
Plesch
Josef
J
Högel
Peter
P
Möller
D000293:Adolescent; D000328:Adult; D000367:Age Factors; D000368:Aged; D015139:Blotting, Southern; D002138:Calibration; D002503:Centromere; D002648:Child; D002675:Child, Preschool; D005260:Female; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D008875:Middle Aged; D012737:Sex Factors; D016615:Telomere / Q000648:ultrastructure*