Abstract

High laser powers are common practice in single molecule localization microscopy (SMLM) 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, multi-color and 3D SMLM with greatly improved resolution and effective labeling efficiency.

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