Abstract

Labelfree nanoscopy encompasses optical imaging with a resolution in the 100-nm range using visible wavelengths. Here, we present a labelfree nanoscopy method that combines coherent imaging techniques with waveguide microscopy to realize a super-condenser featuring maximally inclined coherent darkfield illumination with artificially stretched wave vectors due to large refractive indices of the employed Si3N4 waveguide material. We produce the required coherent plane wave illumination for Fourier ptychography over imaging areas 400 μm2 in size via adiabatically tapered single-mode waveguides and tackle the overlap constraints of the Fourier ptychography phase retrieval algorithm two-fold: first, the directionality of the illumination wave vector is changed sequentially via a multiplexed input structure of the waveguide chip layout, and second, the wave vector modulus is shortend via step-wise increases of the illumination light wavelength over the visible spectrum. We test the method in simulations and in experiments and provide details on the underlying image formation theory as well as the reconstruction algorithm. While the generated Fourier ptychography reconstructions are found to be prone to image artefacts, an alternative coherent imaging method, rotating coherent scattering microscopy (ROCS), is found to be more robust against artefacts but with less achievable resolution.

Highlights

  • Conventional nanoscopy, optical microscopy with resolution below 100 nm, is based on fluorescence [1]

  • Synergistic approaches combining the advantages of label specificity from conventional nanoscopy together with ultra-structural context obtained via labelfree nanoscopy, could bring many new insights about cellular functions, especially as, labelfree nanoscopy has the potential of being applied to living cells and cellular systems

  • It is found that the rotating coherent scattering microscopy (ROCS) imaging procedure results in dramatic enhancement of image contrast, and individual clusters of metal-organic framework (MOF) can be discerned

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Summary

Introduction

Conventional nanoscopy, optical microscopy with resolution below 100 nm, is based on fluorescence [1]. Often listed advantages of nanoscopy, especially in comparison to electron microscopy, are the simple sample preparation, live-cell compatibility, and molecular specificity. Though live-cell compatible, the introduction of fluorescent labels onto the molecular structures of interest are in living cells likely to cause both functional and structural aberrations, potentially leading to false conclusions, and is associated with problems like photobleaching and phototoxicity, variable label specificity, imaging- and image reconstruction-related artifacts, and lengthy optimization protocols [2, 3]. Synergistic approaches combining the advantages of label specificity from conventional nanoscopy together with ultra-structural context obtained via labelfree nanoscopy, could bring many new insights about cellular functions, especially as (contrary to correlative light and electron microscopy [4]), labelfree (optical) nanoscopy has the potential of being applied to living cells and cellular systems.

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