Flow cytometric analysis of microparticles.
Cell-derived microparticles (MPs) are increasingly recognized as important cell-to-cell signaling mechanisms and may exhibit important functions in homeostasis but also in pathogenesis. Indeed, MPs are associated with a number of diseases inhibiting their production that protects against pathogenesis. MPs are distinct from exosomes and apoptotic bodies, often exhibiting the membrane proteins of the activated or apoptotic cell from which they are derived. Electron microscopic analyses have shown that MPs are produced by all cell types tested to date, and ELISA-based assays have established that increased numbers of MPs are produced following cell activation. These approaches do not, however, determine the exact number of MPs and distribution of functional proteins on their surface. Flow cytometry represents an obvious approach to analyze MPs, and we present here a method to assess the number and phenotype of MPs by using a conventional flow cytometer. We also present the caveats with this method and describe a new imaging flow cytometry approach that overcomes these limitations.
Cell Analysis Core Facility, Flow Cytometry, La Jolla Infectious Disease Institute, San Diego, CA, USA.
Henri C
HC
van der Heyde
Irene
I
Gramaglia
Valéry
V
Combes
Thaddeus C
TC
George
Georges E
GE
Grau
D000818:Animals; D000906:Antibodies / Q000378:metabolism; D001773:Blood Cells / Q000378:metabolism; D055252:Cell-Derived Microparticles / Q000378:metabolism*; D042783:Endothelial Cells / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D007091:Image Processing, Computer-Assisted; D051379:Mice; D051381:Rats; D013194:Staining and Labeling
D000906:Antibodies; D005456:Fluorescent Dyes