Extracellular vesicles (EVs) are small, sub-micron membrane-bound particles that function in cell-to-cell communication and have potential to be used in diagnostics, therapy, and biological investigations. However, common characterization methods for EVs lack functional information and rely on “bulk” metrics that lack single-EV resolution. Recent work has applied label-free multimodal nonlinear optical microscopy for characterization of EVs via NAD(P)H and FAD autofluorescence. Here, we characterize EVs isolated from urine and serum from human breast cancer surgery patients and breast reduction surgery patients who have no history of breast cancer to examine altered cancer-related metabolic signatures in cancer-associated EVs.
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