Document Detail


Novel method for cell debris removal in the flow cytometric cell cycle analysis using carboxy-fluorescein diacetate succinimidyl ester.
MedLine Citation:
PMID:  16646047     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: Cell cycle analysis with flow cytometry using propidium iodide (PI) can be difficult in some cases because of the cell debris. Here, we introduce debris removal using intranuclear protein staining (DRIPS), a novel method for separating intact nuclei and cell debris to different populations using carboxy-fluorescein diacetate succinimidyl ester (CFSE). METHODS: To study the apoptosis-sensitivity, chicken DT40 B cell lymphoma cell line was gamma irradiated. After the irradiation, the cells were incubated up to 8 h and the stages of the cell cycle were followed with flow cytometry. RESULTS: CFSE staining, done simultaneously with PI, stained the cell debris brighter than intact nuclei and could be excluded from the histogram with a simple gating procedure. The method is reliable and reproducible and can be executed within 15 min. CONCLUSIONS: DRIPS-method greatly enhances the analysis of difficult cell cycle samples.
Authors:
Perttu Terho; Olli Lassila
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Cytometry. Part A : the journal of the International Society for Analytical Cytology     Volume:  69     ISSN:  1552-4922     ISO Abbreviation:  Cytometry A     Publication Date:  2006 Jun 
Date Detail:
Created Date:  2006-05-29     Completed Date:  2006-07-31     Revised Date:  2007-07-24    
Medline Journal Info:
Nlm Unique ID:  101235694     Medline TA:  Cytometry A     Country:  United States    
Other Details:
Languages:  eng     Pagination:  552-4     Citation Subset:  IM    
Copyright Information:
Copyright 2006 International Society for Analytical Cytology.
Affiliation:
Department of Medical Microbiology, University of Turku, Finland. perttu.terho@utu.fi
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Cycle*
Cell Line, Tumor
Cell Nucleus / metabolism
Chickens / metabolism
Flow Cytometry / methods*
Fluoresceins*
Fluorescent Dyes*
Lymphoma, B-Cell
Staining and Labeling / methods*
Succinimides*
Chemical
Reg. No./Substance:
0/5-(6)-carboxyfluorescein diacetate succinimidyl ester; 0/Fluoresceins; 0/Fluorescent Dyes; 0/Succinimides

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


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