Paper record
Imaging Plant Cells by High-Pressure Freezing and Serial block-face scanning electron microscopy.
Methods in molecular biology (Clifton, N.J.) · 1 Jan 2020 · 10.1007/978-1-0716-0767-1_7
Abstract
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.
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