Abstract
Microparticles, also known as microvesicles, found in blood plasma, urine, and most other body fluids, may serve as valuable biomarkers of diseases such as cardiovascular diseases, systemic inflammatory disease, thrombosis, and cancer. Unfortunately, the detection and quantification of microparticles are hampered by the microscopic size of these particles and their relatively low abundance in blood plasma. The use of a combination of microfluidics and atomic force microscopy to detect microparticles in blood plasma circumvents both problems. In this study, capture of a specific subset of microparticles directly from blood plasma on antibody-coated mica surface is demonstrated. The described method excludes isolation and washing steps to prepare microparticles, improves the detection sensitivity, and yields the size distribution of the captured particles. The majority of the captured particles have a size ranging from 30 to 90 nm, which is in good agreement with prior results obtained with microparticles immediately isolated from fresh plasma. Furthermore, the qualitative shape of the size distribution of microparticles is shown not to be affected by high-speed centrifugation or the use of the microfluidic circuit, demonstrating the relative stable nature of microparticles ex vivo.Electronic supplementary materialThe online version of this article (doi:10.1007/s10544-012-9642-y) contains supplementary material, which is available to authorized users.
Highlights
Blood microparticles (MPs), known as microvesicles, are small particles shed from the surface of many cells upon stimulation or apoptosis (Diamant et al 2004)
The mica surface is functionalized with antibodies to produce a hydrophilic mica surface that cannot bind to the PDMS flow cell
Using image quantification software we found in the samples processed by microfluidic system that there is no significant difference in the number of MPs attached on anti-CD41-coated surface (218, 276, 203, and 240 MPs/100 μm2)
Summary
Blood microparticles (MPs), known as microvesicles, are small particles shed from the surface of many cells upon stimulation or apoptosis (Diamant et al 2004). For a long time they were considered as platelet dust (Wolf 1967), but they have been recognized to participate in important biological processes (Cocucci et al 2009). Examples of such processes are surfacemembrane traffic and the horizontal transfer of protein and RNAs among neighboring cells, which are necessary for rapid phenotype adjustments in a variety of conditions (Cocucci et al 2009). Blood MPs have important physiological and pathological roles in blood coagulation, inflammation and tumor progression (Burnier et al 2009; Pap et al 2009). Flow cytometry (FCM) and capture-based assays are commonly used methods to measure the number of MPs, define their origin based on membrane antigen expression, and asses their procoagulant features
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