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.

doi:10.3389/fpls.2017.01505

Title: A Comparative Study of Sample Preparation for Staining and Immunodetection of Plant Cell Walls by Light Microscopy

Journal Article · Tue Aug 29 00:00:00 EDT 2017 · Frontiers in Plant Science

DOI:https://doi.org/10.3389/fpls.2017.01505· OSTI ID:1398511

Verhertbruggen, Yves ^[1]; Walker, Jesse L. ^[2]; Guillon, Fabienne ^[3]; Scheller, Henrik V. ^[4]

Lawrence Berkeley National Lab. (LBNL), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Institut National de la Recherche Agronomique, Nantes (France)
Lawrence Berkeley National Lab. (LBNL), Emeryville, CA (United States); Univ. of California, Berkeley, CA (United States)
Institut National de la Recherche Agronomique, Nantes (France)
Lawrence Berkeley National Lab. (LBNL), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of California, Berkeley, CA (United States)

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.

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Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1398511

Journal Information:: Frontiers in Plant Science, Vol. 8; ISSN 1664-462X

Publisher:: Frontiers Research FoundationCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 19 works

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
60 APPLIED LIFE SCIENCES
cell wall-degrading enzyme
embedding
fluorescence
immunodetection
microscopy
polysaccharide
sectioning
toluidine blue

Title: A Comparative Study of Sample Preparation for Staining and Immunodetection of Plant Cell Walls by Light Microscopy

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