Journal
NATURE COMMUNICATIONS
Volume 8, Issue -, Pages -Publisher
NATURE PORTFOLIO
DOI: 10.1038/ncomms13558
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Funding
- MRC (Next Generation Optical Microscopy Initiative Program) [MR/K015664/1]
- BBSRC [BB/K01563X/1]
- Royal Society University Research Fellowship
- BBSRC (Sparking Impact program)
- Biotechnology and Biological Sciences Research Council [BB/K01563X/1] Funding Source: researchfish
- Medical Research Council [MR/J000647/1, MR/K015664/1] Funding Source: researchfish
- BBSRC [BB/K01563X/1] Funding Source: UKRI
- MRC [MR/K015664/1, MR/J000647/1] Funding Source: UKRI
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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.
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