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Characterizing blood microparticles: technical aspects and challenges.

Shet AS - Vasc Health Risk Manag (2008)

Bottom Line: MPs that arise from the cellular components of blood and the endothelial lining of blood vessels are referred to as blood MPs and by general consensus are small (< or = 1.5 microm), expose the anionic phospholipid (PL) phosphatidylserine (PS) on the outer leaflet of their membrane, and bear surface membrane antigens reflecting their cellular origin.This brief review summarizes the different approaches used by several groups to study blood MPs.The aim of this article is to review the technical aspects of characterizing the morphological and functional properties of blood MPs with emphasis on the preanalytical and analytical variables involved in these studies.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, St. Johns Medical College and Hospital, St. Johns National Academy of Health Sciences, Bangalore, India. arunshet@iphcr.res.in

ABSTRACT
Although long considered to be cellular debris, microparticles (MPs) are more recently considered reflective of cellular stimulation, activation, and degeneration/apoptosis. MPs that arise from the cellular components of blood and the endothelial lining of blood vessels are referred to as blood MPs and by general consensus are small (< or = 1.5 microm), expose the anionic phospholipid (PL) phosphatidylserine (PS) on the outer leaflet of their membrane, and bear surface membrane antigens reflecting their cellular origin. This brief review summarizes the different approaches used by several groups to study blood MPs. The aim of this article is to review the technical aspects of characterizing the morphological and functional properties of blood MPs with emphasis on the preanalytical and analytical variables involved in these studies.

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Immunogold labeling of blood MPs. Blood MPs obtained by ultracentrifugation were incubated with rabbit polyclonal anti-tissue factor antibody (a; 6 nm gold), anti-CD144 antibody (b; 12 nm gold), and anti-CD14 antibody (c; 12 nm gold). No labeling was observed with control antibodies (d, e, and f).Note: Bars = 100 nm.
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f2-vhrm-4-0769: Immunogold labeling of blood MPs. Blood MPs obtained by ultracentrifugation were incubated with rabbit polyclonal anti-tissue factor antibody (a; 6 nm gold), anti-CD144 antibody (b; 12 nm gold), and anti-CD14 antibody (c; 12 nm gold). No labeling was observed with control antibodies (d, e, and f).Note: Bars = 100 nm.

Mentions: In addition to the above techniques, the antigenic characteristics and the membrane composition of blood MPs have been studied using electron microscopy (Heijnen et al 1999; Aras et al 2004) (Figure 1 and Figure 2), confocal microscopy (Combes et al 1999), high performance liquid chromatography (Weerheim et al 2002), capillary electrophoresis (Xiong et al 2003), and mass spectrometry (Jin et al 2005; Miguet et al 2006) with varying degrees of success. Assays that measure the pro/anticoagulant functions of blood MPs are somewhat under-utilized perhaps in part due to the technical challenges in developing standardized and reproducible assays.


Characterizing blood microparticles: technical aspects and challenges.

Shet AS - Vasc Health Risk Manag (2008)

Immunogold labeling of blood MPs. Blood MPs obtained by ultracentrifugation were incubated with rabbit polyclonal anti-tissue factor antibody (a; 6 nm gold), anti-CD144 antibody (b; 12 nm gold), and anti-CD14 antibody (c; 12 nm gold). No labeling was observed with control antibodies (d, e, and f).Note: Bars = 100 nm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2597765&req=5

f2-vhrm-4-0769: Immunogold labeling of blood MPs. Blood MPs obtained by ultracentrifugation were incubated with rabbit polyclonal anti-tissue factor antibody (a; 6 nm gold), anti-CD144 antibody (b; 12 nm gold), and anti-CD14 antibody (c; 12 nm gold). No labeling was observed with control antibodies (d, e, and f).Note: Bars = 100 nm.
Mentions: In addition to the above techniques, the antigenic characteristics and the membrane composition of blood MPs have been studied using electron microscopy (Heijnen et al 1999; Aras et al 2004) (Figure 1 and Figure 2), confocal microscopy (Combes et al 1999), high performance liquid chromatography (Weerheim et al 2002), capillary electrophoresis (Xiong et al 2003), and mass spectrometry (Jin et al 2005; Miguet et al 2006) with varying degrees of success. Assays that measure the pro/anticoagulant functions of blood MPs are somewhat under-utilized perhaps in part due to the technical challenges in developing standardized and reproducible assays.

Bottom Line: MPs that arise from the cellular components of blood and the endothelial lining of blood vessels are referred to as blood MPs and by general consensus are small (< or = 1.5 microm), expose the anionic phospholipid (PL) phosphatidylserine (PS) on the outer leaflet of their membrane, and bear surface membrane antigens reflecting their cellular origin.This brief review summarizes the different approaches used by several groups to study blood MPs.The aim of this article is to review the technical aspects of characterizing the morphological and functional properties of blood MPs with emphasis on the preanalytical and analytical variables involved in these studies.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, St. Johns Medical College and Hospital, St. Johns National Academy of Health Sciences, Bangalore, India. arunshet@iphcr.res.in

ABSTRACT
Although long considered to be cellular debris, microparticles (MPs) are more recently considered reflective of cellular stimulation, activation, and degeneration/apoptosis. MPs that arise from the cellular components of blood and the endothelial lining of blood vessels are referred to as blood MPs and by general consensus are small (< or = 1.5 microm), expose the anionic phospholipid (PL) phosphatidylserine (PS) on the outer leaflet of their membrane, and bear surface membrane antigens reflecting their cellular origin. This brief review summarizes the different approaches used by several groups to study blood MPs. The aim of this article is to review the technical aspects of characterizing the morphological and functional properties of blood MPs with emphasis on the preanalytical and analytical variables involved in these studies.

Show MeSH
Related in: MedlinePlus