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Exact particle-enhanced point-spread function unlocks 3D super-resolution localization microscopy on nanoparticles

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posted on 2023-06-18, 16:01 authored by Teun A. P. M. Huijben, Sarojini Mahajan, Peter Zijlstra, Rodolphe Marie, Kim I. Mortensen
Nanoparticles (NPs) have proven their applicability in biosensing, drug delivery, and photo-thermal 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 fluorophores point-spread function (PSF). This leads to systematic mislocalizations in conventional analyses employing Gaussian PSFs. Here, we address this shortcoming by deriving the first-ever 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.



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