A method for quantitatively measuring a heterogeneity and a purity of extracellular vesicles from a multiple perspective using a device for a multi-perspective characterization and analysis of the extracellular vesicles, through a system incorporating subsystems for the characterization and analysis of the extracellular vesicles circulating freely in body fluids by multiple methods, comprising:
- characterizing extracellular vesicle phenotype with an atomic force microscopy,
- measuring a size of the extracellular vesicles and detecting topographic coordinates of the extracellular vesicles by combining high-resolution topographic images obtained by the atomic force microscope with software methods,
- measuring a dimensional heterogeneity by translating size measurements of the extracellular vesicles to a size distribution plot and integrating statistical methods for a size-based heterogeneity quantification,
- detecting impurities within samples of the extracellular vesicles based on phenotypical differences of the samples using a machine learning and/or artificial intelligence algorithms
- quantifying a cargo content heterogeneity by performing and comparing a single vesicle level cargo analysis of the extracellular vesicles using a tip-enhanced Raman spectroscopy technique in coordination with known topographic coordinates of the vesicular particles,
- detecting the impurities inside the samples of the extracellular vesicle based on molecular structures of the samples during the single vesicle level cargo analysis.