Reduction of charging in protein electron cryomicroscopy

J. Brink, H. Gross, P. Tittmann, M. B. Sherman, W. Chiu

Research output: Contribution to journalArticle

26 Scopus citations

Abstract

Charging causes a loss of resolution in electron cryomicroscopy with biological specimens prepared without a continuous carbon support film. Thin conductive films were deposited onto catalase crystals prepared across holes using ion-beam sputtering and thermal evaporation and evaluated for the effectiveness of charge reduction. Deposits applied by ion-beam sputtering reduced charging but concurrently resulted in structural damage. Coatings applied by thermal evaporation also reduced charging, and preserved the specimen structure beyond 5 Å resolution as judged from electron diffraction patterns and images of glucose-embedded catalase crystals tilted to 45°in the microscope. This study demonstrates for the first time the feasibility of obtaining high-resolution data from unstained, unsupported protein crystals with a conductive surface coating.

Original languageEnglish (US)
Pages (from-to)67-73
Number of pages7
JournalJournal of Microscopy
Volume191
Issue number1
DOIs
StatePublished - Aug 10 1998
Externally publishedYes

Keywords

  • Charging
  • Electron diffraction pattern
  • Electron gun-equipped evaporator
  • High resolution
  • Ion beam coater
  • Spot-scan imaging
  • Surface deposit

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology

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