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NanoJ-SQUIRREL: quantitative mapping and minimisation of super-resolution optical imaging artefacts

Siân Culley, David Albrecht, Caron Jacobs, Pedro Matos Pereira, View ORCID ProfileChristophe Leterrier, Jason Mercer, View ORCID ProfileRicardo Henriques
doi: https://doi.org/10.1101/158279
Siân Culley
1MRC-Laboratory for Molecular Cell Biology. University College London, Gower Street, London, WC1E 6BT, United Kingdom
2Department of Cell and Developmental Biology, University College London, Gower Street, London, WC1E 6BT, United Kingdom
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David Albrecht
1MRC-Laboratory for Molecular Cell Biology. University College London, Gower Street, London, WC1E 6BT, United Kingdom
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Caron Jacobs
1MRC-Laboratory for Molecular Cell Biology. University College London, Gower Street, London, WC1E 6BT, United Kingdom
2Department of Cell and Developmental Biology, University College London, Gower Street, London, WC1E 6BT, United Kingdom
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Pedro Matos Pereira
1MRC-Laboratory for Molecular Cell Biology. University College London, Gower Street, London, WC1E 6BT, United Kingdom
2Department of Cell and Developmental Biology, University College London, Gower Street, London, WC1E 6BT, United Kingdom
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Christophe Leterrier
3Aix Marseille Université, CNRS, NICN UMR7259, 13344 cedex 15, Marseille, France
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  • ORCID record for Christophe Leterrier
Jason Mercer
1MRC-Laboratory for Molecular Cell Biology. University College London, Gower Street, London, WC1E 6BT, United Kingdom
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  • For correspondence: j.mercer@ucl.ac.uk r.henriques@ucl.ac.uk
Ricardo Henriques
1MRC-Laboratory for Molecular Cell Biology. University College London, Gower Street, London, WC1E 6BT, United Kingdom
2Department of Cell and Developmental Biology, University College London, Gower Street, London, WC1E 6BT, United Kingdom
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  • ORCID record for Ricardo Henriques
  • For correspondence: j.mercer@ucl.ac.uk r.henriques@ucl.ac.uk
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Abstract

Most super-resolution microscopy methods depend on steps that contribute to the formation of image artefacts. Here we present NanoJ-SQUIRREL, an ImageJ-based analytical approach providing a quantitative assessment of super-resolution image quality. By comparing diffraction-limited images and super-resolution equivalents of the same focal volume, this approach generates a quantitative map of super-resolution defects, as well as methods for their correction. To illustrate its broad applicability to super-resolution approaches we apply our method to Localization Microscopy, STED and SIM images of a variety of in-cell structures including microtubules, poxviruses, neuronal actin rings and clathrin coated pits. We particularly focus on single-molecule localisation microscopy, where super-resolution reconstructions often feature imperfections not present in the original data. By showing the quantitative evolution of data quality over these varied sample preparation, acquisition and super-resolution methods we display the potential of NanoJ-SQUIRREL to guide optimization of superresolution imaging parameters.

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Posted July 17, 2017.
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NanoJ-SQUIRREL: quantitative mapping and minimisation of super-resolution optical imaging artefacts
Siân Culley, David Albrecht, Caron Jacobs, Pedro Matos Pereira, Christophe Leterrier, Jason Mercer, Ricardo Henriques
bioRxiv 158279; doi: https://doi.org/10.1101/158279
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NanoJ-SQUIRREL: quantitative mapping and minimisation of super-resolution optical imaging artefacts
Siân Culley, David Albrecht, Caron Jacobs, Pedro Matos Pereira, Christophe Leterrier, Jason Mercer, Ricardo Henriques
bioRxiv 158279; doi: https://doi.org/10.1101/158279

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