| Multiparametric cell cycle analysis by automated microscopy. | |
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MedLine Citation:
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PMID: 16844964 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Cell cycle analysis using flow cytometry (FC) to measure cellular DNA content is a common procedure in drug mechanism of action studies. Although this technique lends itself readily to cell lines that grow in suspension, adherent cell cultures must be resuspended in a cumbersome and potentially invasive procedure that normally involves trypsinization and mechanical agitation of monolayer cultures. High-content analysis (HCA), an automated microscopy-based technology, is well suited to analysis of monolayer cell cultures but provides intrinsically less accurate determination of cellular DNA content than does FC and thus is not the method of choice for cell cycle analysis. Using Cellomics's ArrayScan reader, the authors have developed a 4-color multiparametric HCA approach for cell cycle analysis of adherent cells based on detection of DNA content (4,6-diamidino-2-phenylindole [DAPI] fluorescence), together with the known cell cycle markers bromo-2-deoxyuridine (BrdU) incorporation, cyclin B1 expression, and histone H3 (Ser28) phosphorylation within a single cell population. Considering all 4 markers together, a reliable and accurate quantification of cell cycle phases was possible, as compared with flow cytometric analysis. Using this assay, specific cell cycle blocks induced by treatment with thymidine, paclitaxel, or nocodazole as test drugs were easily monitored in adherent cultures of U-2 OS osteosarcoma cells. |
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Authors:
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Fabio Gasparri; Paolo Cappella; Arturo Galvani |
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Publication Detail:
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Type: Journal Article Date: 2006-07-14 |
Journal Detail:
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Title: Journal of biomolecular screening Volume: 11 ISSN: 1087-0571 ISO Abbreviation: J Biomol Screen Publication Date: 2006 Sep |
Date Detail:
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Created Date: 2006-09-14 Completed Date: 2006-12-12 Revised Date: 2011-05-23 |
Medline Journal Info:
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Nlm Unique ID: 9612112 Medline TA: J Biomol Screen Country: United States |
Other Details:
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Languages: eng Pagination: 586-98 Citation Subset: IM |
Affiliation:
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Biology Department, Oncology, Nerviano Medical Sciences, Nerviano, Italy. fabio.gasparri@nervianoms.com |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Automation Bromodeoxyuridine / pharmacology Cell Cycle / physiology* Cell Division / drug effects Cells, Cultured DNA / analysis Flow Cytometry / instrumentation*, methods Humans Microscopy / instrumentation*, methods |
| Chemical | |
Reg. No./Substance:
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59-14-3/Bromodeoxyuridine; 9007-49-2/DNA |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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