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.
CITATION STYLE
Czymmek, K., Sawant, A., Goodman, K., Pennington, J., Pedersen, P., Hoon, M., & Otegui, M. S. (2020). Imaging Plant Cells by High-Pressure Freezing and Serial block-face scanning electron microscopy. In Methods in Molecular Biology (Vol. 2177, pp. 69–81). Humana Press Inc. https://doi.org/10.1007/978-1-0716-0767-1_7
Mendeley helps you to discover research relevant for your work.