PMID- 9667504
OWN - NLM
STAT- MEDLINE
DCOM- 19990330
LR  - 20191102
IS  - 0196-4763 (Print)
IS  - 0196-4763 (Linking)
VI  - 32
IP  - 3
DP  - 1998 Jul 1
TI  - Assessment of telomere length in hematopoietic interphase cells using in situ 
      hybridization and digital fluorescence microscopy.
PG  - 163-9
AB  - Telomeres are G/C-rich repetitive DNA sequences at the end of all eukaryotic 
      chromosomes. The loss of telomeric repeat sequences during cell divisions has 
      been proposed as a possible mechanism for cell senescence. The standard procedure 
      for measurement of telomere length is Southern blot (SB) hybridization with a 
      telomere-specific probe. However, in using this technique no information can be 
      obtained on variation in telomeric fragments due to interchromosomal, 
      intrachromosomal, and intercellular differences. Lansdorp et al. (Hum Mol Genet 
      5:685-691, 1996) developed a method to measure individual telomeres, using in 
      situ hybridization on metaphase chromosomes, employing peptide nucleic acid (PNA) 
      probes and digital fluorescence microscopy. In this paper we describe a method 
      that can be used to assess telomeric length in interphase cells. An algorithm was 
      developed to measure the total intranuclear fluorescence in situ hybridization 
      (FISH) signal, which features accurate correction for the local autofluorescence. 
      Application of this methodology to samples of fetal liver, umbilical cord blood, 
      and adult bone marrow cells showed a gradual decrease of average telomeric 
      length. Southern blot analysis and PNA FISH measurements on chromosomes in the 
      same samples showed similar results. Advantages of interphase measurements 
      include the possibility of studying nonproliferating cells, thus avoiding 
      selection and cell culturing.
FAU - de Pauw, E S
AU  - de Pauw ES
AD  - Department of Molecular Cell Biology, Leiden University Medical Center, The 
      Netherlands. pauw@rullf2.leidenuniv.nl
FAU - Verwoerd, N P
AU  - Verwoerd NP
FAU - Duinkerken, N
AU  - Duinkerken N
FAU - Willemze, R
AU  - Willemze R
FAU - Raap, A K
AU  - Raap AK
FAU - Fibbe, W E
AU  - Fibbe WE
FAU - Tanke, H J
AU  - Tanke HJ
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Cytometry
JT  - Cytometry
JID - 8102328
SB  - IM
MH  - Adult
MH  - Cell Nucleus/genetics
MH  - Cells, Cultured
MH  - Chromosomes, Human/genetics
MH  - Fetus
MH  - Hematopoietic Stem Cells/*chemistry
MH  - Humans
MH  - *In Situ Hybridization, Fluorescence
MH  - Interphase/*genetics
MH  - Metaphase/genetics
MH  - Microscopy, Fluorescence
MH  - Telomere/*genetics
EDAT- 1998/07/17 03:08
MHDA- 2000/06/20 09:00
CRDT- 1998/07/17 03:08
PHST- 1998/07/17 03:08 [pubmed]
PHST- 2000/06/20 09:00 [medline]
PHST- 1998/07/17 03:08 [entrez]
AID - 10.1002/(SICI)1097-0320(19980701)32:3<163::AID-CYTO1>3.0.CO;2-L [pii]
AID - 10.1002/(sici)1097-0320(19980701)32:3<163::aid-cyto1>3.3.co;2-v [doi]
PST - ppublish
SO  - Cytometry. 1998 Jul 1;32(3):163-9. doi: 
      10.1002/(sici)1097-0320(19980701)32:3<163::aid-cyto1>3.3.co;2-v.