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Article: Combined FISH with pan-telomeric PNA and whole chromosome-specific DNA probes to detect complete and incomplete chromosomal exchanges in human lymphocytes

TitleCombined FISH with pan-telomeric PNA and whole chromosome-specific DNA probes to detect complete and incomplete chromosomal exchanges in human lymphocytes
Authors
Issue Date1999
PublisherInforma Healthcare. The Journal's web site is located at http://www.tandf.co.uk/journals/titles/09553002.asp
Citation
International Journal Of Radiation Biology, 1999, v. 75 n. 9, p. 1107-1112 How to Cite?
AbstractPurpose: To combine FISH with pan-telomeric peptide nucleic acid (PNA) and whole chromosome-specific DNA probes to detect complete and incomplete chromosome exchanges in human lymphocytes. Materials and methods: Human lymphocytes were irradiated in vitro with 0.9 Gy low dose-rate (0.019 Gy/h) tritium β-rays. Metaphase spreads were treated with RNase, fixed in 1:3 acetic acid:methanol, and then further treated with KCl, proteinase K and fixed in 4% paraformaldehyde. Slides were denatured, hybridized for 1.5 h with an FITC-labelled telomeric PNA probe, and rehybridized overnight with a spectrum-orange whole-chromosome probe specific for chromosomes 1, 2 and 4. Hybridized spreads were washed with 70% formamide/20 x SSC and counterstained with DAPI. Results: All three pairs of labelled chromosomes together with 92 telomeres were readily visible after hybridization. The whole chromosomes 1, 2 and 4 were painted orange, and all telomeres were stained green. Unpainted chromosomes were counterstained blue. In the observed 680 chromosome aberrations induced by tritium β-rays in human lymphocytes after 52 h of culture, no evidence of telomere addition was detected. Incomplete and hidden complete exchanges and terminal deletions were definitively discriminated. Conclusion: The simultaneous detection of telomeres and specific whole chromosomes allows for the first time accurate analysis of complete and incomplete chromosome exchanges involving painted chromosomes in human lymphocytes.
Persistent Identifierhttp://hdl.handle.net/10722/178259
ISSN
2015 Impact Factor: 1.779
2015 SCImago Journal Rankings: 0.734
References

 

DC FieldValueLanguage
dc.contributor.authorDeng, Wen_US
dc.contributor.authorLucas, JNen_US
dc.date.accessioned2012-12-19T09:44:48Z-
dc.date.available2012-12-19T09:44:48Z-
dc.date.issued1999en_US
dc.identifier.citationInternational Journal Of Radiation Biology, 1999, v. 75 n. 9, p. 1107-1112en_US
dc.identifier.issn0955-3002en_US
dc.identifier.urihttp://hdl.handle.net/10722/178259-
dc.description.abstractPurpose: To combine FISH with pan-telomeric peptide nucleic acid (PNA) and whole chromosome-specific DNA probes to detect complete and incomplete chromosome exchanges in human lymphocytes. Materials and methods: Human lymphocytes were irradiated in vitro with 0.9 Gy low dose-rate (0.019 Gy/h) tritium β-rays. Metaphase spreads were treated with RNase, fixed in 1:3 acetic acid:methanol, and then further treated with KCl, proteinase K and fixed in 4% paraformaldehyde. Slides were denatured, hybridized for 1.5 h with an FITC-labelled telomeric PNA probe, and rehybridized overnight with a spectrum-orange whole-chromosome probe specific for chromosomes 1, 2 and 4. Hybridized spreads were washed with 70% formamide/20 x SSC and counterstained with DAPI. Results: All three pairs of labelled chromosomes together with 92 telomeres were readily visible after hybridization. The whole chromosomes 1, 2 and 4 were painted orange, and all telomeres were stained green. Unpainted chromosomes were counterstained blue. In the observed 680 chromosome aberrations induced by tritium β-rays in human lymphocytes after 52 h of culture, no evidence of telomere addition was detected. Incomplete and hidden complete exchanges and terminal deletions were definitively discriminated. Conclusion: The simultaneous detection of telomeres and specific whole chromosomes allows for the first time accurate analysis of complete and incomplete chromosome exchanges involving painted chromosomes in human lymphocytes.en_US
dc.languageengen_US
dc.publisherInforma Healthcare. The Journal's web site is located at http://www.tandf.co.uk/journals/titles/09553002.aspen_US
dc.relation.ispartofInternational Journal of Radiation Biologyen_US
dc.subject.meshCells, Cultureden_US
dc.subject.meshChromosome Aberrationsen_US
dc.subject.meshDna Probesen_US
dc.subject.meshHumansen_US
dc.subject.meshIn Situ Hybridization, Fluorescenceen_US
dc.subject.meshLymphocytes - Ultrastructureen_US
dc.subject.meshMaleen_US
dc.subject.meshPeptide Nucleic Acidsen_US
dc.subject.meshTelomereen_US
dc.titleCombined FISH with pan-telomeric PNA and whole chromosome-specific DNA probes to detect complete and incomplete chromosomal exchanges in human lymphocytesen_US
dc.typeArticleen_US
dc.identifier.emailDeng, W: wdeng@hkucc.hku.hken_US
dc.identifier.authorityDeng, W=rp01640en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1080/095530099139575en_US
dc.identifier.pmid10528918-
dc.identifier.scopuseid_2-s2.0-0032851159en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0032851159&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume75en_US
dc.identifier.issue9en_US
dc.identifier.spage1107en_US
dc.identifier.epage1112en_US
dc.publisher.placeUnited Kingdomen_US
dc.identifier.scopusauthoridDeng, W=7202223673en_US
dc.identifier.scopusauthoridLucas, JN=7402441937en_US

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