Teun A.P.M. Huijben, Sarojini Mahajan, Masih Fahim, Peter Zijlstra, Rodolphe Marie, Kim I. Mortensen
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Point-Spread Function Deformations Unlock 3D Localization Microscopy on Spherical Nanoparticles
Nanoparticles (NPs) have proven their applicability in biosensing, drug delivery, and photothermal therapy, but their performance depends critically on the distribution and number of functional groups on their surface. When studying surface functionalization using super-resolution microscopy, the NP modifies the fluorophore’s point-spread function (PSF). This leads to systematic mislocalizations in conventional analyses employing Gaussian PSFs. Here, we address this shortcoming by deriving the analytical PSF model for a fluorophore near a spherical NP. Its calculation is four orders of magnitude faster than numerical approaches and thus feasible for direct use in localization algorithms. We fit this model to individual 2D images from DNA-PAINT experiments on DNA-coated gold NPs and demonstrate extraction of the 3D positions of functional groups with <5 nm precision, revealing inhomogeneous surface coverage. Our method is exact, fast, accessible, and poised to become the standard in super-resolution imaging of NPs for biosensing and drug delivery applications.
期刊介绍:
ACS Nano, published monthly, serves as an international forum for comprehensive articles on nanoscience and nanotechnology research at the intersections of chemistry, biology, materials science, physics, and engineering. The journal fosters communication among scientists in these communities, facilitating collaboration, new research opportunities, and advancements through discoveries. ACS Nano covers synthesis, assembly, characterization, theory, and simulation of nanostructures, nanobiotechnology, nanofabrication, methods and tools for nanoscience and nanotechnology, and self- and directed-assembly. Alongside original research articles, it offers thorough reviews, perspectives on cutting-edge research, and discussions envisioning the future of nanoscience and nanotechnology.