| Phenotypic analysis using very small volumes of blood. | |
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MedLine Citation:
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PMID: 20938921 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Analysis of cell-surface phenotype of peripheral blood leukocytes is one of the most common applications of flow cytometry. In mouse research, the small size of the animal limits the amount of blood available. Standard staining methods using lysis of erythrocytes or gradient separation followed by repeated washing involve unavoidable losses of cells that generally limit analysis of blood to terminal methods. Time-course studies, therefore, require sacrifice of groups of mice at each time point. Thus, a method is needed that can be used with much smaller volumes of blood. This will allow serial sampling of the same animal over time, decreasing experimental variability and reducing animal use. The method described here is a no-lyse, no-wash method that uses triggering on a fluorescence parameter. The method allows routine analysis of the phenotype of peripheral blood leukocytes using whole-blood volumes of 20 µl per tube. The data are comparable with values from traditional methods requiring much higher volumes of blood. Due to interference by erythrocytes, light-scatter parameters are not usable with this method. This method has been used for time-course studies of peripheral blood populations in mice lasting as long as four weeks. |
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Authors:
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James L Weaver; Katherine McKinnon; Dori R Germolec |
Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Current protocols in cytometry / editorial board, J. Paul Robinson, managing editor ... [et al.] Volume: Chapter 6 ISSN: 1934-9300 ISO Abbreviation: Curr Protoc Cytom Publication Date: 2010 Oct |
Date Detail:
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Created Date: 2010-10-12 Completed Date: 2011-02-02 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 100899351 Medline TA: Curr Protoc Cytom Country: United States |
Other Details:
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Languages: eng Pagination: Unit 6.30 Citation Subset: IM |
Affiliation:
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Division of Applied Pharmacology Research, Office of Testing and Research, Center for Drug Evaluation and Research, Food and Drug Administration, Silver Spring, Maryland, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Antibodies / chemistry Autoimmunity Calibration Cell Membrane / metabolism* Flow Cytometry / methods Leukocytes / cytology* Light Lymphocytes / cytology* Mice Models, Animal Phenotype Rats Scattering, Radiation Time Factors |
| Chemical | |
Reg. No./Substance:
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0/Antibodies |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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