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Dual-color single molecule localization microscopy on transparent polymer waveguide chips

View ORCID ProfileAnders Kokkvoll Engdahl, View ORCID ProfileSurjendu Bikash Dutta, Stefan Belle, Jasmin Schürstedt, View ORCID ProfileKarolina Szafranska, Peter McCourt, View ORCID ProfileRalf Hellmann, View ORCID ProfileThomas Huser, View ORCID ProfileMark Schüttpelz
doi: https://doi.org/10.1101/2022.11.29.518375
Anders Kokkvoll Engdahl
1Department of Physics, Bielefeld University, 33615 Bielefeld, Germany
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Surjendu Bikash Dutta
1Department of Physics, Bielefeld University, 33615 Bielefeld, Germany
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Stefan Belle
2Applied Laser and Photonics Group, Aschaffenburg University of Applied Sciences,63743 Aschaffenburg, Germany
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Jasmin Schürstedt
1Department of Physics, Bielefeld University, 33615 Bielefeld, Germany
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Karolina Szafranska
3Department of Medical Biology, UiT - The Arctic University of Norway, N-9019 Tromsø, Norway
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Peter McCourt
3Department of Medical Biology, UiT - The Arctic University of Norway, N-9019 Tromsø, Norway
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Ralf Hellmann
2Applied Laser and Photonics Group, Aschaffenburg University of Applied Sciences,63743 Aschaffenburg, Germany
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Thomas Huser
1Department of Physics, Bielefeld University, 33615 Bielefeld, Germany
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Mark Schüttpelz
1Department of Physics, Bielefeld University, 33615 Bielefeld, Germany
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  • For correspondence: schuettp@physik.uni-bielefeld.de
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Abstract

Photonic waveguide chips offer near-field excitation of biological samples, which enables cost-effective, large field-of-view super-resolution microscopy without the need for high numerical aperture (NA) objective lenses. Single molecule localization based super-resolution microscopy that requires high illumination intensities is currently limited to solid state photonic waveguide chips composed of hard-coated, high NA planar waveguides deposited on opaque substrates. These platforms do not permit epi-detection of fluorescence through the substrate, which limits the use of photonic waveguide chips to the upright configuration. Additionally, the detection efficiency is reduced because the majority of the fluorescence emission is directed towards the high refractive index substrate. A low cost waveguide chip based on a polymer core material deposited on common #1.5 coverslips that is easy to produce was recently demonstrated. Here, a platform that is capable of performing single-molecule localization microscopy (SMLM) of biological samples using polymer-based photonic waveguide chips is presented, enabling super-solution microscopy in the inverted microscope configuration. Super-resolved imaging of two different structures of the cytoskeleton in primary liver sinusoidal endothelial cells (LSECs) by two popular SMLM methods, dSTORM and DNA-PAINT, down to 23 nm is demonstrated.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted December 02, 2022.
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Dual-color single molecule localization microscopy on transparent polymer waveguide chips
Anders Kokkvoll Engdahl, Surjendu Bikash Dutta, Stefan Belle, Jasmin Schürstedt, Karolina Szafranska, Peter McCourt, Ralf Hellmann, Thomas Huser, Mark Schüttpelz
bioRxiv 2022.11.29.518375; doi: https://doi.org/10.1101/2022.11.29.518375
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Dual-color single molecule localization microscopy on transparent polymer waveguide chips
Anders Kokkvoll Engdahl, Surjendu Bikash Dutta, Stefan Belle, Jasmin Schürstedt, Karolina Szafranska, Peter McCourt, Ralf Hellmann, Thomas Huser, Mark Schüttpelz
bioRxiv 2022.11.29.518375; doi: https://doi.org/10.1101/2022.11.29.518375

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