Imaging the Plant Cytoskeleton by High-Pressure Freezing and Electron Tomography.

Electron tomography (ET) imaging of high-pressure frozen/freeze-substituted samples provides a unique opportunity to study structural details of organelles and cytoskeletal arrays in plant cells. In this chapter, we discuss approaches for sample preparation by cryofixation at high pressure, freeze substitution, and resin embedding. We also include pipelines for data collection for electron tomography at ambient temperature, tomogram calculation, and segmentation.