Imaging flow cytometry enhances particle detection sensitivity for extracellular vesicle analysis

exosome rnaHuman peripheral blood mononuclear cells were partitioned into five distinct populations using CD45 and side-scatter intensity (SSC). CCD camera acquisition of fluorescent signal conferred increased sensitivity, facilitating the resolution of less common populations such as eosinophils (yellow) and basophils (white), along with neutrophils, monocytes and lymphocytes (orange, green and blue, respectively). APC-AF750, allophycocyanin–Alexa Fluor 750.

Few areas of biomedical research have generated more interest in the past decade than EV biology. Secreted by most, if not all, human cell types in response to activation, injury or apoptosis, EVs include microvesicles, exosomes and apoptotic bodies. Formed intracellularly or shed by budding of the plasma membrane, these tiny membrane-bound packages of enigmatic macromolecules once overlooked as cellular debris are now recognized as key emissaries mitigating cell-cell interactions. In fact, EVs may be the principal means for the transfer of genetic material and signaling molecules among cells in interstitial spaces and via biological fluids such as plasma, cerebrospinal fluid, sputum and milk. This capacity to ferry bioactive molecules among cells and tissues has underscored the potential significance of EVs in fields as diverse as hematology, regenerative medicine, vascular disease, inflammation and oncology. Here, researchers from Millipore describe how Amnis® IFCs provide a uniquely sensitive platform for the direct detection of EVs while reducing sample volume and preparation time.

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