On the freezing and identification of lipid monolayer 2-D arrays for cryoelectron microscopy

Dianne W. Taylor, Deb Kelly, Anchi Cheng, Kenneth A. Taylor

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Lipid monolayers provide a convenient vehicle for the crystallization of biological macromolecules for 3-D electron microscopy. Although numerous examples of 3-D images from 2-D protein arrays have been described from negatively stained specimens, only six structures have been done from frozen-hydrated specimens. We describe here a method that makes high quality frozen-hydrated specimens of lipid monolayer arrays for cryoelectron microscopy. The method uses holey carbon films with patterned holes for monolayer recovery, blotting and plunge freezing to produce thin aqueous films which cover >90% of the available grid area. With this method, even specimens with relatively infrequent crystals can be screened using automated data collection techniques. Though developed for microscopic examination of 2-D arrays, the method may have wider application to the preparation of single particle specimens for 3-D image reconstruction.

Original languageEnglish (US)
Pages (from-to)305-312
Number of pages8
JournalJournal of Structural Biology
Volume160
Issue number3
DOIs
StatePublished - Dec 1 2007

Fingerprint

Cryoelectron Microscopy
Freezing
Lipids
Three-Dimensional Imaging
Protein Array Analysis
Computer-Assisted Image Processing
Crystallization
Electron Microscopy
Carbon

All Science Journal Classification (ASJC) codes

  • Structural Biology

Cite this

Taylor, Dianne W. ; Kelly, Deb ; Cheng, Anchi ; Taylor, Kenneth A. / On the freezing and identification of lipid monolayer 2-D arrays for cryoelectron microscopy. In: Journal of Structural Biology. 2007 ; Vol. 160, No. 3. pp. 305-312.
@article{59cfa9a063814b678b354eb77c0a9477,
title = "On the freezing and identification of lipid monolayer 2-D arrays for cryoelectron microscopy",
abstract = "Lipid monolayers provide a convenient vehicle for the crystallization of biological macromolecules for 3-D electron microscopy. Although numerous examples of 3-D images from 2-D protein arrays have been described from negatively stained specimens, only six structures have been done from frozen-hydrated specimens. We describe here a method that makes high quality frozen-hydrated specimens of lipid monolayer arrays for cryoelectron microscopy. The method uses holey carbon films with patterned holes for monolayer recovery, blotting and plunge freezing to produce thin aqueous films which cover >90{\%} of the available grid area. With this method, even specimens with relatively infrequent crystals can be screened using automated data collection techniques. Though developed for microscopic examination of 2-D arrays, the method may have wider application to the preparation of single particle specimens for 3-D image reconstruction.",
author = "Taylor, {Dianne W.} and Deb Kelly and Anchi Cheng and Taylor, {Kenneth A.}",
year = "2007",
month = "12",
day = "1",
doi = "10.1016/j.jsb.2007.04.011",
language = "English (US)",
volume = "160",
pages = "305--312",
journal = "Journal of Structural Biology",
issn = "1047-8477",
publisher = "Academic Press Inc.",
number = "3",

}

On the freezing and identification of lipid monolayer 2-D arrays for cryoelectron microscopy. / Taylor, Dianne W.; Kelly, Deb; Cheng, Anchi; Taylor, Kenneth A.

In: Journal of Structural Biology, Vol. 160, No. 3, 01.12.2007, p. 305-312.

Research output: Contribution to journalArticle

TY - JOUR

T1 - On the freezing and identification of lipid monolayer 2-D arrays for cryoelectron microscopy

AU - Taylor, Dianne W.

AU - Kelly, Deb

AU - Cheng, Anchi

AU - Taylor, Kenneth A.

PY - 2007/12/1

Y1 - 2007/12/1

N2 - Lipid monolayers provide a convenient vehicle for the crystallization of biological macromolecules for 3-D electron microscopy. Although numerous examples of 3-D images from 2-D protein arrays have been described from negatively stained specimens, only six structures have been done from frozen-hydrated specimens. We describe here a method that makes high quality frozen-hydrated specimens of lipid monolayer arrays for cryoelectron microscopy. The method uses holey carbon films with patterned holes for monolayer recovery, blotting and plunge freezing to produce thin aqueous films which cover >90% of the available grid area. With this method, even specimens with relatively infrequent crystals can be screened using automated data collection techniques. Though developed for microscopic examination of 2-D arrays, the method may have wider application to the preparation of single particle specimens for 3-D image reconstruction.

AB - Lipid monolayers provide a convenient vehicle for the crystallization of biological macromolecules for 3-D electron microscopy. Although numerous examples of 3-D images from 2-D protein arrays have been described from negatively stained specimens, only six structures have been done from frozen-hydrated specimens. We describe here a method that makes high quality frozen-hydrated specimens of lipid monolayer arrays for cryoelectron microscopy. The method uses holey carbon films with patterned holes for monolayer recovery, blotting and plunge freezing to produce thin aqueous films which cover >90% of the available grid area. With this method, even specimens with relatively infrequent crystals can be screened using automated data collection techniques. Though developed for microscopic examination of 2-D arrays, the method may have wider application to the preparation of single particle specimens for 3-D image reconstruction.

UR - http://www.scopus.com/inward/record.url?scp=36048946749&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=36048946749&partnerID=8YFLogxK

U2 - 10.1016/j.jsb.2007.04.011

DO - 10.1016/j.jsb.2007.04.011

M3 - Article

VL - 160

SP - 305

EP - 312

JO - Journal of Structural Biology

JF - Journal of Structural Biology

SN - 1047-8477

IS - 3

ER -