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

Dianne W. Taylor, Deborah F. Kelly, Anchi Cheng, Kenneth A. Taylor

Research output: Contribution to journalArticle

16 Scopus citations

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 2007

All Science Journal Classification (ASJC) codes

  • Structural Biology

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