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Modernization of Golgi staining techniques for high-resolution, 3-dimensional imaging of individual neurons

Analysis of neuronal arborization and connections is a powerful tool in fundamental and clinical neuroscience. Changes in neuronal morphology are central to brain development and plasticity and are associated with numerous diseases. Golgi staining is a classical technique based on a deposition of me...

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Autores principales: Vints, Katlijn, Vandael, Dorien, Baatsen, Pieter, Pavie, Benjamin, Vernaillen, Frank, Corthout, Nikky, Rybakin, Vasily, Munck, Sebastian, Gounko, Natalia V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6333844/
https://www.ncbi.nlm.nih.gov/pubmed/30644431
http://dx.doi.org/10.1038/s41598-018-37377-x
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author Vints, Katlijn
Vandael, Dorien
Baatsen, Pieter
Pavie, Benjamin
Vernaillen, Frank
Corthout, Nikky
Rybakin, Vasily
Munck, Sebastian
Gounko, Natalia V.
author_facet Vints, Katlijn
Vandael, Dorien
Baatsen, Pieter
Pavie, Benjamin
Vernaillen, Frank
Corthout, Nikky
Rybakin, Vasily
Munck, Sebastian
Gounko, Natalia V.
author_sort Vints, Katlijn
collection PubMed
description Analysis of neuronal arborization and connections is a powerful tool in fundamental and clinical neuroscience. Changes in neuronal morphology are central to brain development and plasticity and are associated with numerous diseases. Golgi staining is a classical technique based on a deposition of metal precipitate in a random set of neurons. Despite their versatility, Golgi methods have limitations that largely precluded their use in advanced microscopy. We combined Golgi staining with fluorescent labeling and tissue clearing techniques in an Alzheimer’s disease model. We further applied 3D electron microscopy to visualize entire Golgi-stained neurons, while preserving ultrastructural details of stained cells, optimized Golgi staining for use with block-face scanning electron microscopy, and developed an algorithm for semi-automated neuronal tracing of cells displaying complex staining patterns. Our method will find use in fundamental neuroscience and the study of neuronal morphology in disease.
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spelling pubmed-63338442019-01-17 Modernization of Golgi staining techniques for high-resolution, 3-dimensional imaging of individual neurons Vints, Katlijn Vandael, Dorien Baatsen, Pieter Pavie, Benjamin Vernaillen, Frank Corthout, Nikky Rybakin, Vasily Munck, Sebastian Gounko, Natalia V. Sci Rep Article Analysis of neuronal arborization and connections is a powerful tool in fundamental and clinical neuroscience. Changes in neuronal morphology are central to brain development and plasticity and are associated with numerous diseases. Golgi staining is a classical technique based on a deposition of metal precipitate in a random set of neurons. Despite their versatility, Golgi methods have limitations that largely precluded their use in advanced microscopy. We combined Golgi staining with fluorescent labeling and tissue clearing techniques in an Alzheimer’s disease model. We further applied 3D electron microscopy to visualize entire Golgi-stained neurons, while preserving ultrastructural details of stained cells, optimized Golgi staining for use with block-face scanning electron microscopy, and developed an algorithm for semi-automated neuronal tracing of cells displaying complex staining patterns. Our method will find use in fundamental neuroscience and the study of neuronal morphology in disease. Nature Publishing Group UK 2019-01-15 /pmc/articles/PMC6333844/ /pubmed/30644431 http://dx.doi.org/10.1038/s41598-018-37377-x Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Vints, Katlijn
Vandael, Dorien
Baatsen, Pieter
Pavie, Benjamin
Vernaillen, Frank
Corthout, Nikky
Rybakin, Vasily
Munck, Sebastian
Gounko, Natalia V.
Modernization of Golgi staining techniques for high-resolution, 3-dimensional imaging of individual neurons
title Modernization of Golgi staining techniques for high-resolution, 3-dimensional imaging of individual neurons
title_full Modernization of Golgi staining techniques for high-resolution, 3-dimensional imaging of individual neurons
title_fullStr Modernization of Golgi staining techniques for high-resolution, 3-dimensional imaging of individual neurons
title_full_unstemmed Modernization of Golgi staining techniques for high-resolution, 3-dimensional imaging of individual neurons
title_short Modernization of Golgi staining techniques for high-resolution, 3-dimensional imaging of individual neurons
title_sort modernization of golgi staining techniques for high-resolution, 3-dimensional imaging of individual neurons
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6333844/
https://www.ncbi.nlm.nih.gov/pubmed/30644431
http://dx.doi.org/10.1038/s41598-018-37377-x
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