| Optimization of yeast cell cycle analysis and morphological characterization by multispectral imaging flow cytometry. | |
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MedLine Citation:
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PMID: 18613038 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Budding yeast Saccharoymyces cerevisiae is a powerful model system for analyzing eukaryotic cell cycle regulation. Yeast cell cycle analysis is typically performed by visual analysis or flow cytometry, and both have limitations in the scope and accuracy of data obtained. This study demonstrates how multispectral imaging flow cytometry (MIFC) provides precise quantitation of cell cycle distribution and morphological phenotypes of yeast cells in flow. Cell cycle analysis of wild-type yeast, nap1Delta, and yeast overexpressing NAP1, was performed visually, by flow cytometry and by MIFC. Quantitative morphological analysis employed measurements of cellular length, thickness, and aspect ratio in an algorithm to calculate a novel feature, bud length. MIFC demonstrated reliable quantification of the yeast cell cycle compared to morphological and flow cytometric analyses. By employing this technique, we observed both the G2/M delay and elongated buds previously described in the nap1Delta strain. Using MIFC, we demonstrate that overexpression of NAP1 causes elongated buds yet only a minor disruption in the cell cycle. The different effects of NAP1 expression level on cell cycle and morphology suggests that these phenotypes are independent. Unlike conventional yeast flow cytometry, MIFC generates complete cell cycle profiles and concurrently offers multiple parameters for morphological analysis. |
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Authors:
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Meredith E K Calvert; Joanne A Lannigan; Lucy F Pemberton |
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Publication Detail:
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Type: Evaluation Studies; Journal Article; Research Support, N.I.H., Extramural |
Journal Detail:
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Title: Cytometry. Part A : the journal of the International Society for Analytical Cytology Volume: 73 ISSN: 1552-4930 ISO Abbreviation: Cytometry A Publication Date: 2008 Sep |
Date Detail:
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Created Date: 2008-09-03 Completed Date: 2008-11-18 Revised Date: 2011-09-26 |
Medline Journal Info:
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Nlm Unique ID: 101235694 Medline TA: Cytometry A Country: United States |
Other Details:
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Languages: eng Pagination: 825-33 Citation Subset: IM |
Affiliation:
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Center for Cell Signaling, University of Virginia, Charlottesville, Virginia 22908, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Cell Cycle* Cell Cycle Proteins / metabolism Cytoskeletal Proteins / metabolism Flow Cytometry / methods* Image Cytometry / methods* Nuclear Proteins / metabolism Saccharomyces cerevisiae / cytology*, growth & development, metabolism Saccharomyces cerevisiae Proteins / metabolism |
| Grant Support | |
ID/Acronym/Agency:
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R01 GM065385-06/GM/NIGMS NIH HHS; R01 GM65385/GM/NIGMS NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Cell Cycle Proteins; 0/Cytoskeletal Proteins; 0/NBP1 protein, S cerevisiae; 0/Nuclear Proteins; 0/Saccharomyces cerevisiae Proteins |
| Comments/Corrections | |
Comment In:
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Cytometry A. 2008 Sep;73(9):777-8
[PMID:
18756513
]
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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