Buzzkill: Surface Acoustic Wave Disruption of Extracellular Vesicles

Abstract Extracellular vesicles (EVs) have emerged as information-rich specimens for potential biomarker analysis. However, their small size, compositional heterogeneity, and limited sample availability present significant analytical challenges. In particular, lipidomic characterization of EVs is complicated by membrane robustness, aggregation behavior, and incompatibility with conventional preparation workflows. Herein, we investigate surface acoustic wave (SAW) manipulation as a mechanical, reagent-free strategy for controlled EV handling across labeling and lipid release regimes. By varying SAW device conditions such as applied powers and frequency, we employed fluorescence studies, nanoparticle tracking analysis, and electron microscopy to demonstrate that low-power, low frequency conditions can minimally disrupt EVs, while promoting dye-vesicle co-localization. We also report that at higher powers and frequencies, SAWs can be further adapted to induce vesicle disruption for direct mass spectrometric analysis, yielding reproducible detection of cholesterol as a representative lipid marker at a threshold of 60 MHz. In this work, we demonstrate the use of SAWs as a versatile tool for addressing key setbacks in EV analyses, offering tunable mechanical features to manipulate EVs, with potential as a sample-preparation approach for future EV lipid analysis.