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
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author Diekmann, Robin
Kahnwald, Maurice
Schoenit, Andreas
Deschamps, Joran
Matti, Ulf
Ries, Jonas
author_facet Diekmann, Robin
Kahnwald, Maurice
Schoenit, Andreas
Deschamps, Joran
Matti, Ulf
Ries, Jonas
author_sort Diekmann, Robin
collection PubMed
description 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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spelling pubmed-76103602021-03-18 Optimizing imaging speed and excitation intensity for single molecule localization microscopy Diekmann, Robin Kahnwald, Maurice Schoenit, Andreas Deschamps, Joran Matti, Ulf Ries, Jonas Nat Methods Article 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. 2020-09-01 2020-08-17 /pmc/articles/PMC7610360/ /pubmed/32807954 http://dx.doi.org/10.1038/s41592-020-0918-5 Text en http://www.nature.com/authors/editorial_policies/license.html#termsUsers may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Diekmann, Robin
Kahnwald, Maurice
Schoenit, Andreas
Deschamps, Joran
Matti, Ulf
Ries, Jonas
Optimizing imaging speed and excitation intensity for single molecule localization microscopy
title Optimizing imaging speed and excitation intensity for single molecule localization microscopy
title_full Optimizing imaging speed and excitation intensity for single molecule localization microscopy
title_fullStr Optimizing imaging speed and excitation intensity for single molecule localization microscopy
title_full_unstemmed Optimizing imaging speed and excitation intensity for single molecule localization microscopy
title_short Optimizing imaging speed and excitation intensity for single molecule localization microscopy
title_sort optimizing imaging speed and excitation intensity for single molecule localization microscopy
topic Article
url 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
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