Abstract
Three-dimensional imaging of cells using electron tomography enables analysis of cell structure at unprecedented resolution. The preparation of cells for tomography using rapid freezing followed by freeze-substitution is an essential first step to ensure the optimal preservation of the cell structure for 3D studies. This protocol outlines a method for obtaining well-preserved cells using high-pressure freezing followed by freeze-substitution. We have found that this method is particularly well suited for electron tomography studies and has the added bonus of preserving antigenicity for immuno-electron microscopy. The steps involved in imaging cells and performing tomographic analysis of cellular structures are also outlined.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 212-216 |
| Number of pages | 5 |
| Journal | Cold Spring Harbor Protocols |
| Volume | 2017 |
| Issue number | 3 |
| DOIs | |
| State | Published - Mar 1 2017 |
| Externally published | Yes |
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
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O’Toole, E. T., Giddings, T. H., & Winey, M. (2017). Cryopreparation and electron tomography of yeast cells. Cold Spring Harbor Protocols, 2017(3), 212-216. https://doi.org/10.1101/pdb.prot085589