PMID- 9685479
OWN - NLM
STAT- MEDLINE
DCOM- 19980922
LR  - 20220317
IS  - 0305-1048 (Print)
IS  - 1362-4962 (Electronic)
IS  - 0305-1048 (Linking)
VI  - 26
IP  - 16
DP  - 1998 Aug 15
TI  - Telomere analysis by fluorescence in situ hybridization and flow cytometry.
PG  - 3651-6
AB  - Determination of telomere length is traditionally performed by Southern blotting 
      and densitometry, giving a mean telomere restriction fragment (TRF) value for the 
      total cell population studied. Fluorescence in situ hybridization (FISH) of 
      telomere repeats has been used to calculate telomere length, a method called 
      quantitative (Q)-FISH. We here present a quantitative flow cytometric approach, 
      Q-FISHFCM, for evaluation of telomere length distribution in individual cells 
      based on in situ hybridization using a fluorescein-labeled peptide nucleic acid 
      (PNA) (CCCTAA)3probe and DNA staining with propidium iodide. A simple and rapid 
      protocol with results within 30 h was developed giving high reproducibility. One 
      important feature of the protocol was the use of an internal cell line control, 
      giving an automatic compensation for potential differences in the hybridization 
      steps. This protocol was tested successfully on cell lines and clinical samples 
      from bone marrow, blood, lymph nodes and tonsils. A significant correlation was 
      found between Southern blotting and Q-FISHFCMtelomere length values ( P = 0.002). 
      The mean sub-telomeric DNA length of the tested cell lines and clinical samples 
      was estimated to be 3.2 kbp. With the Q-FISHFCMmethod the fluorescence signal 
      could be determined in different cell cycle phases, indicating that in human 
      cells the vast majority of telomeric DNA is replicated early in S phase.
FAU - Hultdin, M
AU  - Hultdin M
AD  - Department of Pathology, Umea University, S-90187 Umea, Sweden and Department of 
      Immunocytochemistry, DAKO A/S, DK-2600 Glostrup, Denmark.
FAU - Gronlund, E
AU  - Gronlund E
FAU - Norrback, K
AU  - Norrback K
FAU - Eriksson-Lindstrom, E
AU  - Eriksson-Lindstrom E
FAU - Just, T
AU  - Just T
FAU - Roos, G
AU  - Roos G
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Nucleic Acids Res
JT  - Nucleic acids research
JID - 0411011
RN  - 0 (DNA, Neoplasm)
RN  - 0 (Oligonucleotide Probes)
RN  - 36015-30-2 (Propidium)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Base Sequence
MH  - Blotting, Southern
MH  - Bone Marrow Cells/metabolism/ultrastructure
MH  - Cell Cycle
MH  - Cell Line
MH  - DNA/biosynthesis/genetics
MH  - DNA Replication
MH  - DNA, Neoplasm/biosynthesis/genetics
MH  - Flow Cytometry/*methods
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Neoplasms/genetics/metabolism/ultrastructure
MH  - Oligonucleotide Probes/genetics
MH  - Propidium
MH  - S Phase
MH  - Staining and Labeling
MH  - Telomere/*genetics/metabolism
MH  - Tumor Cells, Cultured
PMC - PMC147775
EDAT- 1998/08/01 00:00
MHDA- 1998/08/01 00:01
PMCR- 1998/08/15
CRDT- 1998/08/01 00:00
PHST- 1998/08/01 00:00 [pubmed]
PHST- 1998/08/01 00:01 [medline]
PHST- 1998/08/01 00:00 [entrez]
PHST- 1998/08/15 00:00 [pmc-release]
AID - gkb610 [pii]
AID - 10.1093/nar/26.16.3651 [doi]
PST - ppublish
SO  - Nucleic Acids Res. 1998 Aug 15;26(16):3651-6. doi: 10.1093/nar/26.16.3651.