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

Abstract

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.

Authors:
 [1];  [2];  [3];  [4]
  1. 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)
  2. Lawrence Berkeley National Lab. (LBNL), Emeryville, CA (United States); Univ. of California, Berkeley, CA (United States)
  3. Institut National de la Recherche Agronomique, Nantes (France)
  4. 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)
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1398511
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Accepted Manuscript
Journal Name:
Frontiers in Plant Science
Additional Journal Information:
Journal Volume: 8; Journal ID: ISSN 1664-462X
Publisher:
Frontiers Research Foundation
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; cell wall-degrading enzyme; embedding; fluorescence; immunodetection; microscopy; polysaccharide; sectioning; toluidine blue

Citation Formats

Verhertbruggen, Yves, Walker, Jesse L., Guillon, Fabienne, and Scheller, Henrik V. A Comparative Study of Sample Preparation for Staining and Immunodetection of Plant Cell Walls by Light Microscopy. United States: N. p., 2017. Web. doi:10.3389/fpls.2017.01505.
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. United States. doi:10.3389/fpls.2017.01505.
Verhertbruggen, Yves, Walker, Jesse L., Guillon, Fabienne, and Scheller, Henrik V. Tue . "A Comparative Study of Sample Preparation for Staining and Immunodetection of Plant Cell Walls by Light Microscopy". United States. doi:10.3389/fpls.2017.01505. https://www.osti.gov/servlets/purl/1398511.
@article{osti_1398511,
title = {A Comparative Study of Sample Preparation for Staining and Immunodetection of Plant Cell Walls by Light Microscopy},
author = {Verhertbruggen, Yves and Walker, Jesse L. and Guillon, Fabienne and Scheller, Henrik V.},
abstractNote = {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.},
doi = {10.3389/fpls.2017.01505},
journal = {Frontiers in Plant Science},
number = ,
volume = 8,
place = {United States},
year = {2017},
month = {8}
}

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Works referenced in this record:

The Cooperative Activities of CSLD2, CSLD3, and CSLD5 Are Required for Normal Arabidopsis Development
journal, November 2011

  • Yin, Lan; Verhertbruggen, Yves; Oikawa, Ai
  • Molecular Plant, Vol. 4, Issue 6
  • DOI: 10.1093/mp/ssr026

Differences in glycosyltransferase family 61 accompany variation in seed coat mucilage composition in Plantago spp.
journal, November 2016

  • Phan, Jana L.; Tucker, Matthew R.; Khor, Shi Fang
  • Journal of Experimental Botany, Vol. 67, Issue 22
  • DOI: 10.1093/jxb/erw424

Use of enzyme-gold complexes for the ultrastructural localization of hemicelluloses in the plant cell wall
journal, January 1984


Understanding How the Complex Molecular Architecture of Mannan-degrading Hydrolases Contributes to Plant Cell Wall Degradation
journal, December 2013

  • Zhang, Xiaoyang; Rogowski, Artur; Zhao, Lei
  • Journal of Biological Chemistry, Vol. 289, Issue 4
  • DOI: 10.1074/jbc.M113.527770

A Comprehensive Toolkit of Plant Cell Wall Glycan-Directed Monoclonal Antibodies
journal, April 2010

  • Pattathil, S.; Avci, U.; Baldwin, D.
  • Plant Physiology, Vol. 153, Issue 2, p. 514-525
  • DOI: 10.1104/pp.109.151985

Carbohydrate-binding modules promote the enzymatic deconstruction of intact plant cell walls by targeting and proximity effects
journal, August 2010

  • Herve, C.; Rogowski, A.; Blake, A. W.
  • Proceedings of the National Academy of Sciences, Vol. 107, Issue 34
  • DOI: 10.1073/pnas.1005732107

Family 46 Carbohydrate-binding Modules Contribute to the Enzymatic Hydrolysis of Xyloglucan and β-1,3–1,4-Glucans through Distinct Mechanisms
journal, February 2015

  • Venditto, Immacolata; Najmudin, Shabir; Luís, Ana S.
  • Journal of Biological Chemistry, Vol. 290, Issue 17
  • DOI: 10.1074/jbc.M115.637827

Collenchyma: a versatile mechanical tissue with dynamic cell walls
journal, August 2012


Embedding of Neural Tissue in Agarose or Glyoxyl Agarose for Vibratome Sectioning
journal, January 1993


The cell wall DUF642 At2g41800 (TEB) protein is involved in hypocotyl cell elongation
journal, December 2016


Understanding How Noncatalytic Carbohydrate Binding Modules Can Display Specificity for Xyloglucan
journal, December 2012

  • Luís, Ana S.; Venditto, Immacolata; Temple, Max J.
  • Journal of Biological Chemistry, Vol. 288, Issue 7
  • DOI: 10.1074/jbc.M112.432781

Inactivation of OsIRX10 Leads to Decreased Xylan Content in Rice Culm Cell Walls and Improved Biomass Saccharification
journal, March 2013

  • Chen, Xuewei; Vega-Sánchez, Miguel E.; Verhertbruggen, Yves
  • Molecular Plant, Vol. 6, Issue 2
  • DOI: 10.1093/mp/sss135

Complementarity of medium-throughput in situ RNA hybridization and tissue-specific transcriptomics: case study of Arabidopsis seed development kinetics
journal, April 2016

  • Francoz, Edith; Ranocha, Philippe; Pernot, Clémentine
  • Scientific Reports, Vol. 6, Issue 1
  • DOI: 10.1038/srep24644

Mutations in Multiple XXT Genes of Arabidopsis Reveal the Complexity of Xyloglucan Biosynthesis
journal, June 2012

  • Zabotina, Olga A.; Avci, Utku; Cavalier, David
  • Plant Physiology, Vol. 159, Issue 4
  • DOI: 10.1104/pp.112.198119

Pectic epitopes are differentially distributed in the cell walls of potato (Solanum tuberosum) tubers
journal, October 1999


Pectin esterification is spatially regulated both within cell walls and between developing tissues of root apices
journal, July 1990

  • Knox, J. Paul; Linstead, PaulJ.; King, Janet
  • Planta, Vol. 181, Issue 4
  • DOI: 10.1007/BF00193004

Non-lignified helical cell wall thickenings in root cortical cells of Aspleniaceae (Polypodiales): histology and taxonomical significance
journal, November 2010

  • Leroux, O.; Bagniewska-Zadworna, A.; Rambe, S. K.
  • Annals of Botany, Vol. 107, Issue 2
  • DOI: 10.1093/aob/mcq225

Immunoprofiling reveals unique cell-specific patterns of wall epitopes in the expanding Arabidopsis stem
journal, February 2013

  • Hall, Hardy C.; Cheung, Jingling; Ellis, Brian E.
  • The Plant Journal, Vol. 74, Issue 1
  • DOI: 10.1111/tpj.12111

Temporal and Spatial Distribution of Pectin Epitopes in Differentiating Anthers and Microspores of Fertile and Sterile Sugar Beet
journal, May 2004

  • Majewska-Sawka, Anna; Münster, Agnieszka; Wisniewska, Ewa
  • Plant and Cell Physiology, Vol. 45, Issue 5
  • DOI: 10.1093/pcp/pch066

Distribution of Fucose-Containing Xyloglucans in Cell Walls of the mur1 Mutant of Arabidopsis
journal, March 2003

  • Freshour, Glenn; Bonin, Christopher P.; Reiter, Wolf-Dieter
  • Plant Physiology, Vol. 131, Issue 4
  • DOI: 10.1104/pp.102.016444

Engineering secondary cell wall deposition in plants
journal, November 2012

  • Yang, Fan; Mitra, Prajakta; Zhang, Ling
  • Plant Biotechnology Journal, Vol. 11, Issue 3
  • DOI: 10.1111/pbi.12016

A conserved functional role of pectic polymers in stomatal guard cells from a range of plant species
journal, December 2004


The Deconstruction of Pectic Rhamnogalacturonan I Unmasks the Occurrence of a Novel Arabinogalactan Oligosaccharide Epitope
journal, September 2015

  • Buffetto, Fanny; Cornuault, Valérie; Rydahl, Maja Gro
  • Plant and Cell Physiology
  • DOI: 10.1093/pcp/pcv128

Monoclonal antibodies to rhamnogalacturonan I backbone
journal, March 2010


The Wheat Grain Contains Pectic Domains Exhibiting Specific Spatial and Development-Associated Distribution
journal, February 2014


Developmental complexity of arabinan polysaccharides and their processing in plant cell walls
journal, August 2009


Distribution of pectic epitopes in cell walls of the sugar beet root
journal, May 2005


Side chains of pectic polysaccharides are regulated in relation to cell proliferation and cell differentiation
journal, December 1999


An extended set of monoclonal antibodies to pectic homogalacturonan
journal, September 2009


Biochemical and immunohistochemical analysis of pectic polysaccharides in the cell walls of Arabidopsis mutant QUASIMODO 1 suspension-cultured cells: implications for cell adhesion
journal, November 2005

  • Leboeuf, Edouard; Guillon, Fabienne; Thoiron, Séverine
  • Journal of Experimental Botany, Vol. 56, Issue 422
  • DOI: 10.1093/jxb/eri314

Pectin Metabolism and Assembly in the Cell Wall of the Charophyte Green Alga Penium margaritaceum
journal, March 2014

  • Domozych, David S.; Sørensen, Iben; Popper, Zoë A.
  • Plant Physiology, Vol. 165, Issue 1
  • DOI: 10.1104/pp.114.236257

Pectic homogalacturonan masks abundant sets of xyloglucan epitopes in plant cell walls
journal, January 2008

  • Marcus, Susan E.; Verhertbruggen, Yves; Hervé, Cécile
  • BMC Plant Biology, Vol. 8, Issue 1
  • DOI: 10.1186/1471-2229-8-60

Comparative glycan profiling of Ceratopteris richardii ‘C-Fern’ gametophytes and sporophytes links cell-wall composition to functional specialization
journal, April 2014

  • Eeckhout, Sharon; Leroux, Olivier; Willats, William G. T.
  • Annals of Botany, Vol. 114, Issue 6
  • DOI: 10.1093/aob/mcu039

ARAD proteins associated with pectic Arabinan biosynthesis form complexes when transiently overexpressed in planta
journal, January 2012

  • Harholt, Jesper; Jensen, Jacob Krüger; Verhertbruggen, Yves
  • Planta, Vol. 236, Issue 1, p. 115-128
  • DOI: 10.1007/s00425-012-1592-3

Antibody-based screening of cell wall matrix glycans in ferns reveals taxon, tissue and cell-type specific distribution patterns
journal, January 2015


Pectin structure and biosynthesis
journal, June 2008


Cell Wall Pectic Arabinans Influence the Mechanical Properties of Arabidopsis thaliana Inflorescence Stems and Their Response to Mechanical Stress
journal, June 2013

  • Verhertbruggen, Yves; Marcus, Susan E.; Chen, Jianshe
  • Plant and Cell Physiology, Vol. 54, Issue 8
  • DOI: 10.1093/pcp/pct074

Enhancing Aniline Blue Fluorescent Staining of Cell Wall Structures
journal, January 1978


Restricted access of proteins to mannan polysaccharides in intact plant cell walls: Masking of mannan polysaccharides
journal, August 2010