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Comprehensive single nanoparticle analysis of synthetic drug delivery systems, as well as natural occurring particles such as Extracellular Vesicles (EVs), is still a major challenge in the field, and is necessary to enhance their successful design, screening and study towards translational application. Investigating population heterogeneity is essential for nanoparticles, as their behaviour, characteristics and thus applicability are strongly affected by this, and cannot be resolved with conventional bulk analysis techniques. Here, we present a dedicated platform for comprehensive Single Particle Automated Raman Trapping Analysis (SPARTA). Nanoparticles ranging from synthetic polymer particles to liposomes or EVs can be integrally analysed by SPARTA without any modification, to obtain their size, determine functionalisation and composition, and monitor dynamic reactions occurring on their surface. The single nanoparticle nature of this approach allows highly detailed investigation in particle heterogeneity, resolving particle mixtures and tracking sequential functionalisations and dynamics on the particle surface. By using a Raman solution marker we demonstrated for the first time the capability to size single nanoparticles in a trap solely by Raman scattering, while simultaneously obtaining their compositional information, allowing novel insights in size-composition relationships. In addition, SPARTA can be applied to study in great detail the biochemical profiles of single EVs from cancerous and non-cancerous origin, towards the use of EVs as cancerous biomarkers for diagnosis, disease progression and evaluating therapeutic efficacy. SPARTA has great potential to critically impact fields from nano drug delivery system design to cancer biomarker identification and profiling.
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