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...
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doaj-8f8de1ce756f4a578893aef37fb4e0672020-11-24T22:15:57ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2017-08-01810.3389/fpls.2017.01505260991A Comparative Study of Sample Preparation for Staining and Immunodetection of Plant Cell Walls by Light MicroscopyYves Verhertbruggen0Yves Verhertbruggen1Yves Verhertbruggen2Jesse L. Walker3Jesse L. Walker4Fabienne Guillon5Henrik V. Scheller6Henrik V. Scheller7Henrik V. Scheller8Joint BioEnergy Institute, Lawrence Berkeley National LaboratoryEmeryville, CA, United StatesEnvironmental Genomics and Systems Biology Division, Lawrence Berkeley National LaboratoryBerkeley, CA, United StatesInstitut National de la Recherche Agronomique, UR 1268Nantes, FranceJoint BioEnergy Institute, Lawrence Berkeley National LaboratoryEmeryville, CA, United StatesDepartment of Plant and Microbial Biology, University of California, BerkeleyBerkeley, CA, United StatesInstitut National de la Recherche Agronomique, UR 1268Nantes, FranceJoint BioEnergy Institute, Lawrence Berkeley National LaboratoryEmeryville, CA, United StatesEnvironmental Genomics and Systems Biology Division, Lawrence Berkeley National LaboratoryBerkeley, CA, United StatesDepartment of Plant and Microbial Biology, University of California, BerkeleyBerkeley, CA, United StatesStaining 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.http://journal.frontiersin.org/article/10.3389/fpls.2017.01505/fullcell wall-degrading enzymeembeddingfluorescenceimmunodetectionmicroscopypolysaccharide |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yves Verhertbruggen Yves Verhertbruggen Yves Verhertbruggen Jesse L. Walker Jesse L. Walker Fabienne Guillon Henrik V. Scheller Henrik V. Scheller Henrik V. Scheller |
spellingShingle |
Yves Verhertbruggen Yves Verhertbruggen Yves Verhertbruggen Jesse L. Walker Jesse L. Walker Fabienne Guillon Henrik V. Scheller Henrik V. Scheller Henrik V. Scheller A Comparative Study of Sample Preparation for Staining and Immunodetection of Plant Cell Walls by Light Microscopy Frontiers in Plant Science cell wall-degrading enzyme embedding fluorescence immunodetection microscopy polysaccharide |
author_facet |
Yves Verhertbruggen Yves Verhertbruggen Yves Verhertbruggen Jesse L. Walker Jesse L. Walker Fabienne Guillon Henrik V. Scheller Henrik V. Scheller Henrik V. Scheller |
author_sort |
Yves Verhertbruggen |
title |
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_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_sort |
comparative study of sample preparation for staining and immunodetection of plant cell walls by light microscopy |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Plant Science |
issn |
1664-462X |
publishDate |
2017-08-01 |
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. |
topic |
cell wall-degrading enzyme embedding fluorescence immunodetection microscopy polysaccharide |
url |
http://journal.frontiersin.org/article/10.3389/fpls.2017.01505/full |
work_keys_str_mv |
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