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Optimizing imaging speed and excitation intensity for single-molecule localization microscopy.

Nature methods (2020-08-19)
Robin Diekmann, Maurice Kahnwald, Andreas Schoenit, Joran Deschamps, Ulf Matti, Jonas Ries
RESUMEN

High laser powers are common practice in single-molecule localization microscopy to speed up data acquisition. Here we systematically quantified how excitation intensity influences localization precision and labeling density, the two main factors determining data quality. We found a strong trade-off between imaging speed and quality and present optimized imaging protocols for high-throughput, multicolor and three-dimensional single-molecule localization microscopy with greatly improved resolution and effective labeling efficiency.

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