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Imaging Plant Cells by High-Pressure Freezing and Serial block-face scanning electron microscopy

Methods in Molecular Biology, ISSN: 1940-6029, Vol: 2177, Page: 69-81
2020
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Book Chapter Description

This chapter describes methods to enhanced contrast of plant material processed by high-pressure freezing and freeze substitution for improved visualization by serial block-face scanning electron microscopy (SBEM). The contrast enhancing steps are based on a protocol involving the sequential incubation of samples in heavy metals and sodium thiocarbohydrazide (OTO staining). We also describe the pipeline for imaging plant tissues in a commercial SBEM system (Gatan 3View) and routines for the image analysis and three-dimensional reconstructions using open-source and commercial software packages.

Bibliographic Details

Czymmek, Kirk; Sawant, Abhilash; Goodman, Kaija; Pennington, Janice; Pedersen, Pal; Hoon, Mrinalini; Otegui, Marisa S

Springer Science and Business Media LLC

Biochemistry, Genetics and Molecular Biology

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