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Local dimensionality determines imaging speed in localization microscopy

Author

Listed:
  • Patrick Fox-Roberts

    (King’s College London)

  • Richard Marsh

    (King’s College London)

  • Karin Pfisterer

    (King’s College London)

  • Asier Jayo

    (King’s College London)

  • Maddy Parsons

    (King’s College London)

  • Susan Cox

    (King’s College London)

Abstract

Localization microscopy allows biological samples to be imaged at a length scale of tens of nanometres. Live-cell super-resolution imaging is rare, as it is generally assumed to be too slow for dynamic samples. The speed of data acquisition can be optimized by tuning the density of activated fluorophores in each time frame. Here, we show that the maximum achievable imaging speed for a particular structure varies by orders of magnitude, depending on the sample dimensionality (that is, whether the sample is more like a point, a strand or an extended structure such as a focal adhesion). If too high an excitation density is used, we demonstrate that the analysis undergoes silent failure, resulting in reconstruction artefacts. We are releasing a tool to allow users to identify areas of the image in which the activation density was too high and correct for them, in both live- and fixed-cell experiments.

Suggested Citation

  • Patrick Fox-Roberts & Richard Marsh & Karin Pfisterer & Asier Jayo & Maddy Parsons & Susan Cox, 2017. "Local dimensionality determines imaging speed in localization microscopy," Nature Communications, Nature, vol. 8(1), pages 1-10, April.
    • Handle: RePEc:nat:natcom:v:8:y:2017:i:1:d:10.1038_ncomms13558
    DOI: 10.1038/ncomms13558
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