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bioRxiv · 10.1101/2025.03.13.643103

Super-resolving particle diffusion heterogeneity in porous hydrogels via high-speed 3D active-feedback single-particle tracking microscopy

Lin, Y.·Lin, H.·Welsher, K.
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Abstract

Nanoparticle diffusion in 3D porous structures is critical to understanding natural and synthetic systems but remains underexplored due to limitations in traditional microscopy methods. Here, we use 3D Single-Molecule Active-feedback Real-time Tracking (3D-SMART) microscopy to resolve nanoparticle dynamics in agarose gels with unprecedented spatiotemporal resolution. We highlight hopping diffusion, where particles intermittently escape confinement pockets, providing insights into hydrogel microstructure. Long, highly sampled trajectories enable extraction of kinetic parameters, confinement sizes, and thermodynamic barriers. This study demonstrates 3D-SMARTs ability to probe particle-environment interactions at super-resolution ([~]10 nm in XY and [~]30 nm in Z) in 3D, offering new perspectives on nanoparticle diffusion and the structural dynamics of porous materials, with implications for drug delivery, material science, and biological systems. ToC figure O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=182 SRC="FIGDIR/small/643103v3_ufig1.gif" ALT="Figure 1"> View larger version (45K): org.highwire.dtl.DTLVardef@7c11c4org.highwire.dtl.DTLVardef@1725c22org.highwire.dtl.DTLVardef@fecc4forg.highwire.dtl.DTLVardef@175d33_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Lin, Y., Lin, H., Welsher, K.. 2025-03-15. Super-resolving particle diffusion heterogeneity in porous hydrogels via high-speed 3D active-feedback single-particle tracking microscopy. https://doi.org/10.1101/2025.03.13.643103

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