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 |
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ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
Cite this
Cryopreparation and electron tomography of yeast cells. / O’Toole, Eileen T.; Giddings, Thomas H.; Winey, Mark.
In: Cold Spring Harbor Protocols, Vol. 2017, No. 3, 01.03.2017, p. 212-216.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Cryopreparation and electron tomography of yeast cells
AU - O’Toole, Eileen T.
AU - Giddings, Thomas H.
AU - Winey, Mark
PY - 2017/3/1
Y1 - 2017/3/1
N2 - 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.
AB - 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.
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U2 - 10.1101/pdb.prot085589
DO - 10.1101/pdb.prot085589
M3 - Article
VL - 2017
SP - 212
EP - 216
JO - Cold Spring Harbor Protocols
JF - Cold Spring Harbor Protocols
SN - 1559-6095
IS - 3
ER -