High-Resolution Field Emission Scanning Electron Microscopy (FESEM) Imaging of Cellulose Microfibril Organization in Plant Primary Cell Walls

doi:10.1017/S143192761701251X

Title: High-Resolution Field Emission Scanning Electron Microscopy (FESEM) Imaging of Cellulose Microfibril Organization in Plant Primary Cell Walls

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Abstract

Abstract We have used field emission scanning electron microscopy (FESEM) to study the high-resolution organization of cellulose microfibrils in onion epidermal cell walls. We frequently found that conventional “rule of thumb” conditions for imaging of biological samples did not yield high-resolution images of cellulose organization and often resulted in artifacts or distortions of cell wall structure. Here we detail our method of one-step fixation and dehydration with 100% ethanol, followed by critical point drying, ultrathin iridium (Ir) sputter coating (3 s), and FESEM imaging at a moderate accelerating voltage (10 kV) with an In-lens detector. We compare results obtained with our improved protocol with images obtained with samples processed by conventional aldehyde fixation, graded dehydration, sputter coating with Au, Au/Pd, or carbon, and low-voltage FESEM imaging. The results demonstrated that our protocol is simpler, causes little artifact, and is more suitable for high-resolution imaging of cell wall cellulose microfibrils whereas such imaging is very challenging by conventional methods.

Authors:: Zheng, Yunzhen; Cosgrove, Daniel J.; Ning, Gang

Publication Date:: 2017-08-24

Research Org.:: Energy Frontier Research Centers (EFRC) (United States). Center for Lignocellulose Structure and Formation (CLSF)

Sponsoring Org.:: USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)

OSTI Identifier:: 1469867

DOE Contract Number:: SC0001090

Resource Type:: Journal Article

Journal Name:: Microscopy and Microanalysis

Additional Journal Information:: Journal Volume: 23; Journal Issue: 05; Related Information: CLSF partners with Pennsylvania State University (lead); North Carolina State University; University of Rhode Island; Virginia Tech University; Journal ID: ISSN 1431-9276

Publisher:: Microscopy Society of America (MSA)

Country of Publication:: United States

Language:: English

Subject:: biofuels (including algae and biomass), bio-inspired, membrane, carbon sequestration, materials and chemistry by design, synthesis (self-assembly)

Citation Formats


                    Zheng, Yunzhen, Cosgrove, Daniel J., and Ning, Gang. High-Resolution Field Emission Scanning Electron Microscopy (FESEM) Imaging of Cellulose Microfibril Organization in Plant Primary Cell Walls.  United States: N. p., 2017. 
        Web.  doi:10.1017/S143192761701251X.

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                    Zheng, Yunzhen, Cosgrove, Daniel J., & Ning, Gang. High-Resolution Field Emission Scanning Electron Microscopy (FESEM) Imaging of Cellulose Microfibril Organization in Plant Primary Cell Walls.  United States.  doi:10.1017/S143192761701251X.

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                    Zheng, Yunzhen, Cosgrove, Daniel J., and Ning, Gang. Thu .  
        "High-Resolution Field Emission Scanning Electron Microscopy (FESEM) Imaging of Cellulose Microfibril Organization in Plant Primary Cell Walls".  United States.  doi:10.1017/S143192761701251X.

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@article{osti_1469867,

  title        = {High-Resolution Field Emission Scanning Electron Microscopy (FESEM) Imaging of Cellulose Microfibril Organization in Plant Primary Cell Walls},

  author       = {Zheng, Yunzhen and Cosgrove, Daniel J. and Ning, Gang},

  abstractNote = {Abstract We have used field emission scanning electron microscopy (FESEM) to study the high-resolution organization of cellulose microfibrils in onion epidermal cell walls. We frequently found that conventional “rule of thumb” conditions for imaging of biological samples did not yield high-resolution images of cellulose organization and often resulted in artifacts or distortions of cell wall structure. Here we detail our method of one-step fixation and dehydration with 100% ethanol, followed by critical point drying, ultrathin iridium (Ir) sputter coating (3 s), and FESEM imaging at a moderate accelerating voltage (10 kV) with an In-lens detector. We compare results obtained with our improved protocol with images obtained with samples processed by conventional aldehyde fixation, graded dehydration, sputter coating with Au, Au/Pd, or carbon, and low-voltage FESEM imaging. The results demonstrated that our protocol is simpler, causes little artifact, and is more suitable for high-resolution imaging of cell wall cellulose microfibrils whereas such imaging is very challenging by conventional methods.},

  doi          = {10.1017/S143192761701251X},

  journal      = {Microscopy and Microanalysis},

  issn         = {1431-9276},

  number       = 05,

  volume       = 23,

  place        = {United States},

  year         = {2017},

  month        = {8}

}

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DOI: 10.1017/S143192761701251X

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