Table 1 Summary of some methods applied for MPs research
Method | Quantification | Cell origin and/or function identification | MPs size distribution | Limitations | References |
|---|---|---|---|---|---|
Electron microscopy | Limited | Limited (only for single labeling by immunoelectron microscopy) | Yes, but might be subjective due to limited number of measurements | Artifacts due to specimen preparation for negative contrast (drying, application of contrasting solution etc.) | Hess et al., 1999; Distler et al., 2005; Lima et al., 2009; Witek et al., 2009; Porro et al., 2010; Duarte et al., 2012; Gercel-Taylor et al., 2012 |
Functional assays (procoagulant activity, thrombin generation tests, ELISA-based tests etc.) | Yes (bulk) | No | No | Only information on procoagulant or thrombin generating activity available | Leroyer et al., 2007; Tesselaar et al., 2007; Salzer et al., 2008; Manly et al., 2009; Van der Heyde et al., 2011 |
Atomic Force Microscopy | Limited | Limited (requires development of AB-coated surfaces) | Yes, but might be subjective due to limited number of measurements | Artifacts due to abundance of cell debri and plasma protein | Salzer et al., 2008; Yuana et al., 2010; Leong et al., 2011; Nantakomol et al., 2012 |
Light scattering techniques (nanoparticle tracking analysis, submicron particle analysis, dynamic light scattering) | Yes | No* | Yes | Artifacts due to abundance of cell debri and plasma protein – samples requires special purification | Lawrie et al., 2009; Xu et al., 2010; Gercel-Taylor et al., 2012 |
Western blotting | Semi-quantitative | Yes | No | Requires significant amount of starting material (> 10 μg of vesicular material) | Abid Hussein et al., 2005; Salzer et al., 2008; Sander et al., 2008; Bebawy et al., 2009; Bernimoulin et al., 2009; Gercel-Taylor et al., 2012 |
Mass-spectrometry | No | Yes, allows identification of multiple proteins | No | Requires significant amount of starting material | Sander et al., 2008; Mayr et al., 2009; Rood et al., 2010 |
Flow Cytometry | Yes | Yes, allows identification of multiple antigens | Limited | Limited; >300 nm particle range (conventional flow cytometry); presence of protein aggregates may lead to artifacts sensitivity depends on cytometer | Orozco, Lewis, 2010; Zwicker et al., 2010; Ayers et al., 2011; Yuana et al., 2011; van der Heyde et al., 2011 |
Flow imaging cytometry | Yes | Yes, allows quantification of multiple antigens | No | Limited for bright fluorescence MPs | Van der Heyde et al., 2011 |