| Cell-cycle synchronisation of bloodstream forms of Trypanosoma brucei using Vybrant DyeCycle Violet-based sorting. | |
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MedLine Citation:
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PMID: 19729042 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Studies on the cell-cycle of Trypanosoma brucei have revealed several unusual characteristics that differ from the model eukaryotic organisms. However, the inability to isolate homogenous populations of parasites in distinct cell-cycle stages has limited the analysis of trypanosome cell division and complicated the understanding of mutant phenotypes with possible impact on cell-cycle related events. Although hydroxyurea-induced cell-cycle arrest in procyclic and bloodstream forms has been applied recently with success, such block-release protocols can complicate the analysis of cell-cycle regulated events and have the potential to disrupt important cell-cycle checkpoints. An alternative approach based on flow cytometry of parasites stained with Vybrant DyeCycle Orange circumvents this problem, but is restricted to procyclic form parasites. Here, we apply Vybrant Dyecycle Violet staining coupled with flow cytometry to effectively select different cell-cycle stages of bloodstream form trypanosomes. Moreover, the sorted parasites remain viable, although synchrony is rapidly lost. This method enables cell-cycle enrichment of populations of trypanosomes in their mammal infective stage, particularly at the G1 phase. |
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Authors:
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Sarah Kabani; Martin Waterfall; Keith R Matthews |
Publication Detail:
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Type: Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't Date: 2009-09-01 |
Journal Detail:
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Title: Molecular and biochemical parasitology Volume: 169 ISSN: 1872-9428 ISO Abbreviation: Mol. Biochem. Parasitol. Publication Date: 2010 Jan |
Date Detail:
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Created Date: 2009-11-16 Completed Date: 2010-01-25 Revised Date: 2010-09-24 |
Medline Journal Info:
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Nlm Unique ID: 8006324 Medline TA: Mol Biochem Parasitol Country: Netherlands |
Other Details:
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Languages: eng Pagination: 59-62 Citation Subset: IM |
Affiliation:
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Centre for Immunity, Infection and Evolution, Institute of Immunology and Infection Research, University of Edinburgh, United Kingdom. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Cell Cycle* Cell Line Flow Cytometry / methods* Fluorescent Dyes / chemistry Staining and Labeling Trypanosoma brucei brucei / chemistry*, cytology* Trypanosomiasis, African / parasitology* |
| Grant Support | |
ID/Acronym/Agency:
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073358//Wellcome Trust; //Biotechnology and Biological Sciences Research Council; //Wellcome Trust |
| Chemical | |
Reg. No./Substance:
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0/Fluorescent Dyes |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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