Document Detail


Interactions of intercalating fluorochromes with DNA analyzed by conventional and fluorescence lifetime flow cytometry utilizing deuterium oxide.
MedLine Citation:
PMID:  8891446     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Deuterium oxide (D2O) has been shown in previous studies to increase both the fluorescence lifetime and fluorescence intensity of propidium iodide (PI) and ethidium bromide (EB) when bound to nucleic acid structures. We have used spectroscopic analysis and conventional and phase-sensitive flow cytometry to compare changes in PI and EB fluorescence intensity and lifetime bound to DNA and fixed Chinese hamster ovary (CHO) cells in the presence of D2O vs. phosphate-buffered saline (PBS). Spectroscopic and flow cytometric studies showed a twofold enhancement of fluorescence intensity of PI and EB bound to fixed CHO cells in D2O and a 5 ns increase in PI and EB fluorescence lifetimes in D2O. The fluorescence lifetime of HL-60 cells stained with PI or EB was found to be 1-2 ns different from that of CHO cells, indicating that the lifetime of these fluorochromes is sensitive to chromatin configuration in different cells types. Apoptotic subpopulations of HL-60 cells had a significantly reduced fluorescence lifetime compared to nonapoptotic subpopulations. Results indicate that different chromatin states, or differences in the structures of PI and EB, lead to alterations in the fluorescence intensity and fluorescence lifetime of these intercalating probes.
Authors:
B L Sailer; A J Nastasi; J G Valdez; J A Steinkamp; H A Crissman
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Cytometry     Volume:  25     ISSN:  0196-4763     ISO Abbreviation:  Cytometry     Publication Date:  1996 Oct 
Date Detail:
Created Date:  1997-01-24     Completed Date:  1997-01-24     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  8102328     Medline TA:  Cytometry     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  164-72     Citation Subset:  IM    
Affiliation:
Life Sciences Division, Los Alamos National Laboratory, NM 87545, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Apoptosis
CHO Cells
Cattle
Cricetinae
DNA / drug effects*
Deuterium Oxide*
Ethidium
Flow Cytometry*
Fluorescent Dyes / pharmacology*
HL-60 Cells
Humans
Intercalating Agents / pharmacology*
Propidium
Sodium Chloride
Spectrometry, Fluorescence
Grant Support
ID/Acronym/Agency:
P41-RR01315/RR/NCRR NIH HHS; R01 RR06758/RR/NCRR NIH HHS; R01 RR07855/RR/NCRR NIH HHS
Chemical
Reg. No./Substance:
0/Fluorescent Dyes; 0/Intercalating Agents; 3546-21-2/Ethidium; 36015-30-2/Propidium; 7647-14-5/Sodium Chloride; 7789-20-0/Deuterium Oxide; 9007-49-2/DNA

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


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