Automated cryoEM data acquisition and analysis of 284742 particles of GroEL.

Automated cryoEM data acquisition and analysis of 284742 particles of GroEL. Research Abstract Details 

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Automated cryoEM data acquisition and analysis of 284742 particles of GroEL. Abstract Text:

Scott M Stagg,Gabriel C Lander,James Pulokas,Denis Fellmann,Anchi Cheng,Joel D Quispe,Satya P Mallick,Radomir M Avila,Bridget Carragher,Clinton S Potter,

One of the goals in developing our automated electron microscopy data acquisition system, Leginon, was to improve both the ease of use and the throughput of the process of acquiring low dose images of macromolecular specimens embedded in vitreous ice. In this article, we demonstrate the potential of the Leginon system for high-throughput data acquisition by describing an experiment in which we acquired images of more than 280,000 particles of GroEL in a single 25 h session at the microscope. We also demonstrate the potential for an automated pipeline for molecular microscopy by showing that these particles can be subjected to completely automated procedures to reconstruct a three-dimensional (3D) density map to a resolution better than 8 A. In generating the 3D maps, we used a variety of metadata associated with the data acquisition and processing steps to sort and select the particles. These metadata provide a number of insights into factors that affect the quality of the acquired images and the resulting reconstructions. In particular, we show that the resolution of the reconstructed 3D density maps improves with decreasing ice thickness. These data provide a basis for assessing the capabilities of high-throughput macromolecular microscopy.

Automated cryoEM data acquisition and analysis of 284742 particles of GroEL. Publishing Authors By Initials

SM Stagg,GC Lander,J Pulokas,D Fellmann,A Cheng,JD Quispe,SP Mallick,RM Avila,B Carragher,CS Potter,

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Automated cryoEM data acquisition and analysis of 284742 particles of GroEL. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Journal of structural biology

VOLUME: 155

Page Numbers: 470-81

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ISSN: 1047-8477

DAY: 22

MONTH: 05

YEAR: 2006

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LANGUAGE: eng

NlmUniqueID: 9011206

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Grant and Affiliation Information for Automated cryoEM data acquisition and analysis of 284742 particles of GroEL.

AFFILIATION: The National Resource for Automated Molecular Microscopy, Department of Cell Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.

Country: United States

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MEDLINETA: J Struct Biol

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