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A Comparative Study of Sample Preparation for Staining and Immunodetection of Plant Cell Walls by Light Microscopy
Staining and immunodetection by light microscopy are methods widely used to investigate plant cell walls. The two techniques have been crucial to study the cell wall architecture in planta, its deconstruction by chemicals or cell wall-degrading enzymes. They have been instrumental in detecting the p...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5581911/ https://www.ncbi.nlm.nih.gov/pubmed/28900439 http://dx.doi.org/10.3389/fpls.2017.01505 |
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author | Verhertbruggen, Yves Walker, Jesse L. Guillon, Fabienne Scheller, Henrik V. |
author_facet | Verhertbruggen, Yves Walker, Jesse L. Guillon, Fabienne Scheller, Henrik V. |
author_sort | Verhertbruggen, Yves |
collection | PubMed |
description | Staining and immunodetection by light microscopy are methods widely used to investigate plant cell walls. The two techniques have been crucial to study the cell wall architecture in planta, its deconstruction by chemicals or cell wall-degrading enzymes. They have been instrumental in detecting the presence of cell types, in deciphering plant cell wall evolution and in characterizing plant mutants and transformants. The success of immunolabeling relies on how plant materials are embedded and sectioned. Agarose coating, wax and resin embedding are, respectively, associated with vibratome, microtome and ultramicrotome sectioning. Here, we have systematically carried out a comparative analysis of these three methods of sample preparation when they are applied for cell wall staining and cell wall immunomicroscopy. In order to help the plant community in understanding and selecting adequate methods of embedding and sectioning for cell wall immunodetection, we review in this article the advantages and limitations of these three methods. Moreover, we offer detailed protocols of embedding for studying plant materials through microscopy. |
format | Online Article Text |
id | pubmed-5581911 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-55819112017-09-12 A Comparative Study of Sample Preparation for Staining and Immunodetection of Plant Cell Walls by Light Microscopy Verhertbruggen, Yves Walker, Jesse L. Guillon, Fabienne Scheller, Henrik V. Front Plant Sci Plant Science Staining and immunodetection by light microscopy are methods widely used to investigate plant cell walls. The two techniques have been crucial to study the cell wall architecture in planta, its deconstruction by chemicals or cell wall-degrading enzymes. They have been instrumental in detecting the presence of cell types, in deciphering plant cell wall evolution and in characterizing plant mutants and transformants. The success of immunolabeling relies on how plant materials are embedded and sectioned. Agarose coating, wax and resin embedding are, respectively, associated with vibratome, microtome and ultramicrotome sectioning. Here, we have systematically carried out a comparative analysis of these three methods of sample preparation when they are applied for cell wall staining and cell wall immunomicroscopy. In order to help the plant community in understanding and selecting adequate methods of embedding and sectioning for cell wall immunodetection, we review in this article the advantages and limitations of these three methods. Moreover, we offer detailed protocols of embedding for studying plant materials through microscopy. Frontiers Media S.A. 2017-08-29 /pmc/articles/PMC5581911/ /pubmed/28900439 http://dx.doi.org/10.3389/fpls.2017.01505 Text en Copyright © 2017 Verhertbruggen, Walker, Guillon and Scheller. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Plant Science Verhertbruggen, Yves Walker, Jesse L. Guillon, Fabienne Scheller, Henrik V. A Comparative Study of Sample Preparation for Staining and Immunodetection of Plant Cell Walls by Light Microscopy |
title | A Comparative Study of Sample Preparation for Staining and Immunodetection of Plant Cell Walls by Light Microscopy |
title_full | A Comparative Study of Sample Preparation for Staining and Immunodetection of Plant Cell Walls by Light Microscopy |
title_fullStr | A Comparative Study of Sample Preparation for Staining and Immunodetection of Plant Cell Walls by Light Microscopy |
title_full_unstemmed | A Comparative Study of Sample Preparation for Staining and Immunodetection of Plant Cell Walls by Light Microscopy |
title_short | A Comparative Study of Sample Preparation for Staining and Immunodetection of Plant Cell Walls by Light Microscopy |
title_sort | comparative study of sample preparation for staining and immunodetection of plant cell walls by light microscopy |
topic | Plant Science |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5581911/ https://www.ncbi.nlm.nih.gov/pubmed/28900439 http://dx.doi.org/10.3389/fpls.2017.01505 |
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