Confounds in Functional Magnetic Resonance Imaging-Part I: Mechanisms.

Extracellular vesicles (EV) are becoming crucial targets in liquid biopsy, diagnostics, and therapeutic applications, yet their nanoscale characterization remains challenging. In this context, the detection of drug-induced liver injury, i.e., hepatotoxicity, through EV molecular content remains unexplored. To this end, we present the SPECTRA - Surface Plasmonic Enhancement with Combined Transformative RAman and Interferometric Microscopy approach, which provides rapid, label-free EV content analysis under 30 minutes and requires only 1.3 microliters of sample. Using hepatic cultures as a model, our platform captures distinct and reproducible EV molecular changes in response to acetaminophen-induced hepatotoxicity. Across independent culture sets, a Gaussian Process Regression model predicted Acetaminophen (APAP) dose with median absolute error of 1.50 mM and minimal bias (Bland-Altman bias ~0.03 mM); cross-validated RMSE was 3.145 mM. These findings establish EVs as dynamic reporters of cellular drug responses and demonstrate use of SPECTRA for EV detection of hepatotoxicity.