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Tailoring cryo-electron microscopy grids by photo-micropatterning for in-cell structural studies

Spatially-controlled cell adhesion on electron microscopy (EM) supports remains a bottleneck in specimen preparation for cellular cryo-electron tomography. Here, we describe contactless and mask-free photo-micropatterning of EM grids for site-specific deposition of extracellular matrix (ECM)-related...

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Autores principales: Toro-Nahuelpan, Mauricio, Zagoriy, Ievgeniia, Senger, Fabrice, Blanchoin, Laurent, Théry, Manuel, Mahamid, Julia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6949126/
https://www.ncbi.nlm.nih.gov/pubmed/31740821
http://dx.doi.org/10.1038/s41592-019-0630-5
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author Toro-Nahuelpan, Mauricio
Zagoriy, Ievgeniia
Senger, Fabrice
Blanchoin, Laurent
Théry, Manuel
Mahamid, Julia
author_facet Toro-Nahuelpan, Mauricio
Zagoriy, Ievgeniia
Senger, Fabrice
Blanchoin, Laurent
Théry, Manuel
Mahamid, Julia
author_sort Toro-Nahuelpan, Mauricio
collection PubMed
description Spatially-controlled cell adhesion on electron microscopy (EM) supports remains a bottleneck in specimen preparation for cellular cryo-electron tomography. Here, we describe contactless and mask-free photo-micropatterning of EM grids for site-specific deposition of extracellular matrix (ECM)-related proteins. We attained refined cell positioning for micromachining by cryo-focused ion beam milling. Complex micropatterns generated predictable intracellular organization, allowing direct correlation between cell architecture and in-cell 3D-structural characterization of the underlying molecular machinery.
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spelling pubmed-69491262020-05-18 Tailoring cryo-electron microscopy grids by photo-micropatterning for in-cell structural studies Toro-Nahuelpan, Mauricio Zagoriy, Ievgeniia Senger, Fabrice Blanchoin, Laurent Théry, Manuel Mahamid, Julia Nat Methods Article Spatially-controlled cell adhesion on electron microscopy (EM) supports remains a bottleneck in specimen preparation for cellular cryo-electron tomography. Here, we describe contactless and mask-free photo-micropatterning of EM grids for site-specific deposition of extracellular matrix (ECM)-related proteins. We attained refined cell positioning for micromachining by cryo-focused ion beam milling. Complex micropatterns generated predictable intracellular organization, allowing direct correlation between cell architecture and in-cell 3D-structural characterization of the underlying molecular machinery. 2020-01-01 2019-11-18 /pmc/articles/PMC6949126/ /pubmed/31740821 http://dx.doi.org/10.1038/s41592-019-0630-5 Text en http://www.nature.com/authors/editorial_policies/license.html#terms Users 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
Toro-Nahuelpan, Mauricio
Zagoriy, Ievgeniia
Senger, Fabrice
Blanchoin, Laurent
Théry, Manuel
Mahamid, Julia
Tailoring cryo-electron microscopy grids by photo-micropatterning for in-cell structural studies
title Tailoring cryo-electron microscopy grids by photo-micropatterning for in-cell structural studies
title_full Tailoring cryo-electron microscopy grids by photo-micropatterning for in-cell structural studies
title_fullStr Tailoring cryo-electron microscopy grids by photo-micropatterning for in-cell structural studies
title_full_unstemmed Tailoring cryo-electron microscopy grids by photo-micropatterning for in-cell structural studies
title_short Tailoring cryo-electron microscopy grids by photo-micropatterning for in-cell structural studies
title_sort tailoring cryo-electron microscopy grids by photo-micropatterning for in-cell structural studies
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6949126/
https://www.ncbi.nlm.nih.gov/pubmed/31740821
http://dx.doi.org/10.1038/s41592-019-0630-5
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