Optimizing imaging speed and excitation intensity for single molecule localization microscopy

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 t...

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Detalles Bibliográficos
Autores principales: Diekmann, Robin, Kahnwald, Maurice, Schoenit, Andreas, Deschamps, Joran, Matti, Ulf, Ries, Jonas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2020
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7610360/
https://www.ncbi.nlm.nih.gov/pubmed/32807954
http://dx.doi.org/10.1038/s41592-020-0918-5
Descripción
Sumario: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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