Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 114, Issue 35, Pages 9273-9278Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1704908114
Keywords
superresolution microscopy; 3D reconstruction; DNA origami; multivariate statistical analysis; structural biology
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Funding
- European Research Council (ERC) Starting Grant [ERC-Stg-260787]
- French National Research Agency [ANR-16-CE09-0004-01, ANR-15-CE09-0003-02]
- BioMedical Engineering startup fund
- Comision Nacional de Investigacion Cientifica y Tecnologica, Chile, PAI/INDUSTRIA [79090016]
- France-BioImaging [ANR-10-INSB-04]
- Agence Nationale de la Recherche (ANR) [ANR-16-CE09-0004] Funding Source: Agence Nationale de la Recherche (ANR)
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Superresolution light microscopy allows the imaging of labeled supramolecular assemblies at a resolution surpassing the classical diffraction limit. A serious limitation of the superresolution approach is sample heterogeneity and the stochastic character of the labeling procedure. To increase the reproducibility and the resolution of the superresolution results, we apply multivariate statistical analysis methods and 3D reconstruction approaches originally developed for cryogenic electron microscopy of single particles. These methods allow for the reference-free 3D reconstruction of nanomolecular structures from two-dimensional superresolution projection images. Since these 2D projection images all show the structure in high-resolution directions of the optical microscope, the resulting 3D reconstructions have the best possible isotropic resolution in all directions.
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