Abstract
Inefficient delivery of macromolecules and nanoparticles to intracellular targets is a major bottleneck in drug delivery, genetic engineering, and molecular imaging. Here we apply live-cell single-quantum-dot imaging and tracking to analyze and classify nanoparticle states after intracellular delivery. By merging trajectory diffusion parameters with brightness measurements, multidimensional analysis reveals distinct and heterogeneous populations that are indistinguishable using single parameters alone. We derive new quantitative metrics of particle loading, cluster distribution, and vesicular release in single cells, and evaluate intracellular nanoparticles with diverse surfaces following osmotic delivery. Surface properties have a major impact on cell uptake, but little impact on the absolute cytoplasmic numbers. A key outcome is that stable zwitterionic surfaces yield uniform cytosolic behavior, ideal for imaging agents. We anticipate that this combination of quantum dots and single-particle tracking can be widely applied to design and optimize next-generation imaging probes, nanoparticle therapeutics, and biologics.
Highlights
Inefficient delivery of macromolecules and nanoparticles to intracellular targets is a major bottleneck in drug delivery, genetic engineering, and molecular imaging
quantum dots (QDs) were coated with five different polymeric coatings, depicted schematically in Fig. 1d, with hydrodynamic diameter (h.d.) and electrostatic charge
Fmobile for pPEG-QDs increased substantially when delivered through liposomal vesicles, reaching 78% (Supplementary Fig. 16), demonstrating that adsorption processes occurring during delivery can have a dominating impact on the intracellular state
Summary
Inefficient delivery of macromolecules and nanoparticles to intracellular targets is a major bottleneck in drug delivery, genetic engineering, and molecular imaging. We apply live-cell single-quantum-dot imaging and tracking to analyze and classify nanoparticle states after intracellular delivery. We derive new quantitative metrics of particle loading, cluster distribution, and vesicular release in single cells, and evaluate intracellular nanoparticles with diverse surfaces following osmotic delivery. A key outcome is that stable zwitterionic surfaces yield uniform cytosolic behavior, ideal for imaging agents We anticipate that this combination of quantum dots and singleparticle tracking can be widely applied to design and optimize next-generation imaging probes, nanoparticle therapeutics, and biologics. We apply live-cell single-nanoparticle fluorescence imaging and tracking to quantitatively evaluate nanoparticle state distributions following intracellular delivery. We analyze intracellular QD trajectories to derive new classification metrics that distinguish distinct intracellular states that have previously been inaccessible through ensemble methods with the goal of mechanistically evaluating intracellular delivery of nanoparticles and macromolecules. QDs with zwitterionic surfaces are the most mobile and homogeneously dispersed after delivery, adding to the rapidly expanding utility of zwitterionic nanomaterials
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