Document Detail


Quantitative fluorescence in situ hybridization (QFISH) of telomere lengths in tissue and cells.
MedLine Citation:
PMID:  18770817     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Telomeres are repetitive DNA sequences at the end of each chromosome that provide stability and prevent end-to-end chromosome fusions. In order to understand mechanisms responsible for telomere shortening, it is necessary to develop methods for accurate telomere length measurement that can be applied to archival and fresh tissue and cells. This unit describes in situ-based quantitative fluorescence in situ hybridization (QFISH) protocols using a fluorescence-conjugated telomere probe (peptide nucleic acid, PNA) that stains telomeres proportionally to their length. These protocols can be used on formalin-fixed paraffin-embedded tissue, lightly fixed tissue, cells isolated from tissue, cultured cells, and agar-embedded cells. The basic protocol for QFISH staining is modified to achieve excellent QFISH staining for a variety of cell preparations. Image-analysis techniques to quantitate average telomere lengths from tissues and isolated stained cells are also described.
Authors:
Jacintha N O'Sullivan; Jennifer C Finley; Rosa-Ana Risques; Wen-Tang Shen; Katherine A Gollahon; Peter S Rabinovitch
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Current protocols in cytometry / editorial board, J. Paul Robinson, managing editor ... [et al.]     Volume:  Chapter 12     ISSN:  1934-9300     ISO Abbreviation:  Curr Protoc Cytom     Publication Date:  2005 Aug 
Date Detail:
Created Date:  2008-09-04     Completed Date:  2008-10-08     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  100899351     Medline TA:  Curr Protoc Cytom     Country:  United States    
Other Details:
Languages:  eng     Pagination:  Unit 12.6     Citation Subset:  IM    
Affiliation:
St Vincent's University Hospital, Elm Park, Dublin, Ireland.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cells / cytology,  ultrastructure
Chromosomes / physiology,  ultrastructure
Formaldehyde
Image Processing, Computer-Assisted
In Situ Hybridization / methods*
Spectrometry, Fluorescence / methods
Telomere / ultrastructure*
Chemical
Reg. No./Substance:
50-00-0/Formaldehyde

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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