Reduction of charging in protein electron cryomicroscopy

J. Brink, H. Gross, P. Tittmann, Michael Sherman, W. Chiu

Research output: Contribution to journalArticle

25 Citations (Scopus)

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 - 1998
Externally publishedYes

Fingerprint

Cryoelectron Microscopy
Catalase
charging
catalase
Thermal evaporation
Hot Temperature
Ions
proteins
Proteins
Ion beams
Crystals
Sputtering
Electrons
sputtering
ion beams
evaporation
crystals
coatings
Coatings
Conductive films

Keywords

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

ASJC Scopus subject areas

  • Instrumentation

Cite this

Reduction of charging in protein electron cryomicroscopy. / Brink, J.; Gross, H.; Tittmann, P.; Sherman, Michael; Chiu, W.

In: Journal of Microscopy, Vol. 191, No. 1, 1998, p. 67-73.

Research output: Contribution to journalArticle

Brink, J. ; Gross, H. ; Tittmann, P. ; Sherman, Michael ; Chiu, W. / Reduction of charging in protein electron cryomicroscopy. In: Journal of Microscopy. 1998 ; Vol. 191, No. 1. pp. 67-73.
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