Local dimensionality determines imaging speed in localization microscopy

Patrick Fox-Roberts, Richard Marsh, Karin Pfisterer, Asier Jayo, Maddy Parsons and Susan Cox ()
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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

Nature Communications, 2017, vol. 8, issue 1, 1-10

Abstract: 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.

Date: 2017
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DOI: 10.1038/ncomms13558

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