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

Part of: Micrographia Collection

Published online by Cambridge University Press:  24 August 2017

Yunzhen Zheng ,
Daniel J. Cosgrove  and
Gang Ning
Yunzhen Zheng
Affiliation:
Department of Biology, Penn State University, University Park, PA 16802, USA
Daniel J. Cosgrove
Affiliation:
Department of Biology, Penn State University, University Park, PA 16802, USA
Gang Ning*
Affiliation:
Department of Biology, Penn State University, University Park, PA 16802, USA Huck Institutes of the Life Sciences, Penn State University, University Park, PA 16802, USA
*
*Corresponding author. [email protected]
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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.

Keywords

high-resolution scanning electron microscopyfield emission scanning electron microscopyepidermal cell wallscellulose microfibrilsiridium sputter coating
Type
Micrographia
Information
Microscopy and Microanalysis , Volume 23 , Issue 5 , October 2017 , pp. 1048 - 1054
Copyright
© Microscopy Society of America 2017 

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