Scaling structure factor amplitudes in electron cryomicroscopy using X- ray solution scattering

Michael F. Schmid, Michael B. Sherman, Paul Matsudaira, Hiro Tsuruta, Wah Chiu

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

The structure factors derived from electron cryomicroscopic images are modified by the contrast transfer function of the microscope's objective lens and other influences. The phases of the structure factors can be corrected in a straightforward way when the positions of the contrast transfer function rings are determined. However, corrected amplitudes are also essential to yield an accurate distribution of mass in the reconstruction. The correct scale factors for the amplitudes are difficult to evaluate for data that are merged from many different micrographs. We opt to use X-ray solution scattering intensity from a concentrated suspension of the specimen to correct the amplitudes of the spherically averaged structure factors. When this approach is applied to the three-dimensional image data of ice-embedded acrosomal bundles, the core of a filament in a three-dimensional reconstruction of the acrosomal bundle becomes denser and matches more closely the outer density ascribed to scruin.

Original languageEnglish (US)
Pages (from-to)51-57
Number of pages7
JournalJournal of Structural Biology
Volume128
Issue number1
DOIs
StatePublished - Dec 1 1999
Externally publishedYes

ASJC Scopus subject areas

  • Structural Biology

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