Large-volume en-bloc staining for electron microscopy-based connectomics

Large-scale connectomics requires dense staining of neuronal tissue blocks for electron microscopy (EM). Here we report a large-volume dense en-bloc EM staining protocol that overcomes the staining gradients, which so far substantially limited the reconstructable volumes in three-dimensional (3D) EM...

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Detalles Bibliográficos
Autores principales: Hua, Yunfeng, Laserstein, Philip, Helmstaedter, Moritz
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Pub. Group 2015
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4532871/
https://www.ncbi.nlm.nih.gov/pubmed/26235643
http://dx.doi.org/10.1038/ncomms8923
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author Hua, Yunfeng
Laserstein, Philip
Helmstaedter, Moritz
author_facet Hua, Yunfeng
Laserstein, Philip
Helmstaedter, Moritz
author_sort Hua, Yunfeng
collection PubMed
description Large-scale connectomics requires dense staining of neuronal tissue blocks for electron microscopy (EM). Here we report a large-volume dense en-bloc EM staining protocol that overcomes the staining gradients, which so far substantially limited the reconstructable volumes in three-dimensional (3D) EM. Our protocol provides densely reconstructable tissue blocks from mouse neocortex sized at least 1 mm in diameter. By relaxing the constraints on precise topographic sample targeting, it makes the correlated functional and structural analysis of neuronal circuits realistic.
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spelling pubmed-45328712015-08-31 Large-volume en-bloc staining for electron microscopy-based connectomics Hua, Yunfeng Laserstein, Philip Helmstaedter, Moritz Nat Commun Article Large-scale connectomics requires dense staining of neuronal tissue blocks for electron microscopy (EM). Here we report a large-volume dense en-bloc EM staining protocol that overcomes the staining gradients, which so far substantially limited the reconstructable volumes in three-dimensional (3D) EM. Our protocol provides densely reconstructable tissue blocks from mouse neocortex sized at least 1 mm in diameter. By relaxing the constraints on precise topographic sample targeting, it makes the correlated functional and structural analysis of neuronal circuits realistic. Nature Pub. Group 2015-08-03 /pmc/articles/PMC4532871/ /pubmed/26235643 http://dx.doi.org/10.1038/ncomms8923 Text en Copyright © 2015, Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved. http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Hua, Yunfeng
Laserstein, Philip
Helmstaedter, Moritz
Large-volume en-bloc staining for electron microscopy-based connectomics
title Large-volume en-bloc staining for electron microscopy-based connectomics
title_full Large-volume en-bloc staining for electron microscopy-based connectomics
title_fullStr Large-volume en-bloc staining for electron microscopy-based connectomics
title_full_unstemmed Large-volume en-bloc staining for electron microscopy-based connectomics
title_short Large-volume en-bloc staining for electron microscopy-based connectomics
title_sort large-volume en-bloc staining for electron microscopy-based connectomics
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4532871/
https://www.ncbi.nlm.nih.gov/pubmed/26235643
http://dx.doi.org/10.1038/ncomms8923
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AT helmstaedtermoritz largevolumeenblocstainingforelectronmicroscopybasedconnectomics

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