Electron Tomography and Immunogold Labelling as Tools to Analyse De Novo Assembly of Plant Cell Walls
High-resolution imaging of the membranous intermediates and cytoskeletal arrays involved in the assembly of a new cell wall during plant cytokinesis requires state-of-the-art electron microscopy techniques. The combination of cryofixation/freeze-substitution methods with electron tomography (ET) has revealed amazing structural details of this unique cellular process. This chapter deals with the main steps associated with these imaging techniques: selection of samples suitable for studying plant cytokinesis, sample preparation by high-pressure freezing/freeze substitution, and ET of plastic sections. In addition, immunogold approaches for the identification of proteins and polysaccharides during cell wall assembly are discussed.
Key wordsElectron tomography Cell walls Cytokinesis Cryofixation Immunolabelling
This work was supported by the National Research Initiative of the USDA Cooperative State Research, Education and Extension Service, grant number 2008-35304-18672 to M.S.O.
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