Cryopreparation and electron tomography of yeast cells

Eileen T. O’Toole; Thomas H. Giddings; Mark Winey

doi:10.1101/pdb.prot085589

Cryopreparation and electron tomography of yeast cells

Eileen T. O’Toole, Thomas H. Giddings, Mark Winey

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

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	https://doi.org/10.1101/pdb.prot085589
State	Published - Mar 1 2017
Externally published	Yes

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)

Access to Document

10.1101/pdb.prot085589

Fingerprint Dive into the research topics of 'Cryopreparation and electron tomography of yeast cells'. Together they form a unique fingerprint.

Cite this

@article{41acff73edcb406d94ec774e1e9e35bb,

title = "Cryopreparation and electron tomography of yeast cells",

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.",

author = "O{\textquoteright}Toole, {Eileen T.} and Giddings, {Thomas H.} and Mark Winey",

year = "2017",

month = mar,

day = "1",

doi = "10.1101/pdb.prot085589",

language = "English (US)",

volume = "2017",

pages = "212--216",

journal = "Cold Spring Harbor Protocols",

issn = "1559-6095",

publisher = "Cold Spring Harbor Laboratory Press",

number = "3",

}

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.

UR - http://www.scopus.com/inward/record.url?scp=85014685376&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85014685376&partnerID=8YFLogxK

U2 - 10.1101/pdb.prot085589

DO - 10.1101/pdb.prot085589

M3 - Article

C2 - 28250212

AN - SCOPUS:85014685376

VL - 2017

SP - 212

EP - 216

JO - Cold Spring Harbor Protocols

JF - Cold Spring Harbor Protocols

SN - 1559-6095

IS - 3

ER -

Cryopreparation and electron tomography of yeast cells

Abstract

ASJC Scopus subject areas

Access to Document

Other files and links

Cite this