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Flat-field illumination for quantitative fluorescence imaging

Ian Khaw, Benjamin Croop, Jialei Tang, Anna Möhl, Ulrike Fuchs, View ORCID ProfileKyu Young Han
doi: https://doi.org/10.1101/297242
Ian Khaw
1CREOL, The College of Optics and Photonics, University of Central Florida, Orlando, Florida, USA.
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Benjamin Croop
1CREOL, The College of Optics and Photonics, University of Central Florida, Orlando, Florida, USA.
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Jialei Tang
1CREOL, The College of Optics and Photonics, University of Central Florida, Orlando, Florida, USA.
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Anna Möhl
2asphericon GmbH, Stockholmer Str. 9, D-07745 Jena, Germany
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Ulrike Fuchs
2asphericon GmbH, Stockholmer Str. 9, D-07745 Jena, Germany
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Kyu Young Han
1CREOL, The College of Optics and Photonics, University of Central Florida, Orlando, Florida, USA.
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  • ORCID record for Kyu Young Han
  • For correspondence: kyhan@creol.ucf.edu
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Abstract

The uneven illumination of a Gaussian profile makes quantitative analysis highly challenging in laser-based wide-field fluorescence microscopy. Here we present flat-field illumination (FFI) where the Gaussian beam is reshaped into a uniform flat-top profile using a high-precision refractive optical component. The long working distance and high spatial coherence of FFI allows us to accomplish uniform epi and TIRF illumination for multi-color single-molecule imaging. In addition, high-throughput borderless imaging is demonstrated with minimal image overlap.

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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-ND 4.0 International license.
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Posted April 11, 2018.
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Flat-field illumination for quantitative fluorescence imaging
Ian Khaw, Benjamin Croop, Jialei Tang, Anna Möhl, Ulrike Fuchs, Kyu Young Han
bioRxiv 297242; doi: https://doi.org/10.1101/297242
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Flat-field illumination for quantitative fluorescence imaging
Ian Khaw, Benjamin Croop, Jialei Tang, Anna Möhl, Ulrike Fuchs, Kyu Young Han
bioRxiv 297242; doi: https://doi.org/10.1101/297242

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