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Characterization of Nanoparticles in Suspension by Simultaneous iNTA and Fluorescence Detection with Single-Molecule Sensitivity

Jiang, S.; Kashkanova, A. D.; Lee, H.; Miller, M. E. C.; Utikal, T.; Shkarin, A.; Qazvini, H.; Sandoghdar, V.

2026-04-13 biophysics
10.64898/2026.04.09.717488 bioRxiv
Show abstract

Quantitative knowledge of nanoparticle properties is desirable in a large number of scientific and technological applications, but measurements with a high degree of precision usually prove to be challenging. Among a range of available methodologies, optical techniques with single particle sensitivity are especially interesting because they can reveal intrinsic hetero-geneities in a fast non-invasive manner. Recently, we presented interferometric nanoparticle tracking analysis (iNTA) as a highly sensitive label-free technique that is capable of determining the size, concentration and index of refraction of different subpopulations in a suspension mixture. Here, we enhance this method with biochemical specificity through multicolor fluorescence detection at the single-molecule sensitivity limit. We benchmark the performance of the combined technique, which we name iNTA-F, by distinguishing populations of fluorescent and non-fluorescent nanoparticles of different material, size, and fluorescence intensity, with an emphasis on the characterization of lipid vesicles and biological extracellular vesicles (EVs).

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