| Flow cytometric analysis of circulating microparticles in plasma. | |
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MedLine Citation:
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PMID: 20235276 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Microparticles, which include exosomes, micro-vesicles, apoptotic bodies and apoptotic microparticles, are small (0.05 - 3 mum in diameter), membranous vesicles that can contain DNA, RNA, miRNA, intracellular proteins and express extracellular surface markers from the parental cells. They can be secreted from intracellular multivesicular bodies or released from the surface of blebbing membranes. Circulating microparticles are abundant in the plasma of normal individuals and can be derived from circulating blood cells such as platelets, red blood cells and leukocytes as well as from tissue sources, such as endothelial and placental tissues. Elevated levels of microparticles are associated with various diseases such as thrombosis (platelet microparticles), congestive heart failure (endothelial microparticles), breast cancer patients (leukocyte microparticles) and women with preeclampsia (syncytiotrophoblast microparticles). Although microparticles can be detected by microscopy, enzyme-linked immunoassays and functional assays, flow cytometry is the preferred method because of the ability to quantitate (fluorescent bead- or flow rate-based method) and because of polychromatic capabilities. However, standardization of pre-analytical and analytical modus operandi for isolating, enumerating and fluorescent labeling of microparticles remains a challenge. The primary focus of this article is to review the preliminary steps required to optimally study circulating in vivo microparticles which include: 1) centrifugation speed used, 2) quantitation of microparticles before antibody labeling, 3) levels of fluorescence intensity of antibody-labeled microparticles, 4) polychromatic flow cytometric analysis of microparticle sub-populations and 5) use of polyclonal antibodies designed for Western blotting for flow cytometry. These studies determine a roadmap to develop microparticles as biomarkers for a variety of conditions. |
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Authors:
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Aaron F Orozco; Dorothy E Lewis |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Review |
Journal Detail:
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Title: Cytometry. Part A : the journal of the International Society for Analytical Cytology Volume: 77 ISSN: 1552-4930 ISO Abbreviation: Cytometry A Publication Date: 2010 Jun |
Date Detail:
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Created Date: 2010-05-27 Completed Date: 2010-09-15 Revised Date: 2011-07-28 |
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: 502-14 Citation Subset: IM |
Copyright Information:
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Copyright 2010 International Society for Advancement of Cytometry. |
Affiliation:
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University of Texas Health Science Center at Houston, Internal Medicine/Infectious Diseases, Houston, Texas 77030, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Blood Chemical Analysis
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methods* Blotting, Western Cell Line Cell-Derived Microparticles / chemistry* Flow Cytometry* Fluorescence Humans Staining and Labeling |
| Grant Support | |
ID/Acronym/Agency:
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HDO46623//PHS HHS; R01 HD046623-03/HD/NICHD NIH HHS; T32AI007495/AI/NIAID NIH HHS |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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