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TitleRevisiting sodium phosphotungstate and ammonium molybdate as nonradioactive negative-staining agents for single-particle analysis.
Journal, issue, pagesActa Crystallogr F Struct Biol Commun, Vol. 80, Issue Pt 12, Page 356-362, Year 2024
Publish dateDec 1, 2024
AuthorsMonika Gunkel / Arthur Macha / Elmar Behrmann /
PubMed AbstractThis study reports the successful replacement of uranyl-based stains by either sodium phosphotungstate or ammonium molybdate in negative-staining electron microscopy. Using apoferritin as a test ...This study reports the successful replacement of uranyl-based stains by either sodium phosphotungstate or ammonium molybdate in negative-staining electron microscopy. Using apoferritin as a test specimen, it is demonstrated that in combination with a facile on-grid fixation step, both stains yield comparable images to uranyl formate. Subsequently, using β-galactosidase, it is shown that both stains can also successfully be employed for single-particle analysis, yielding virtually indistinguishable results from uranyl formate. As both replacement stains are nonradioactive, they are not subjected to the same handling restrictions as uranyl-based stains. Therefore they are not only cheaper to use, but also make decentralized sample-grid preparation, directly after purification, accessible to a broader range of scientists.
External linksActa Crystallogr F Struct Biol Commun / PubMed:39601624 / PubMed Central
MethodsEM (single particle)
Resolution9.5 - 10.06 Å
Structure data

EMDB-51069: UF-stained beta-galactosidase tetramer
Method: EM (single particle) / Resolution: 9.5 Å

EMDB-51071: SPT-stained beta-galactosidase tetramer
Method: EM (single particle) / Resolution: 9.6 Å

EMDB-51072: AMo stained beta-galactosidase
Method: EM (single particle) / Resolution: 10.06 Å

Source
  • Escherichia coli (E. coli)
  • Escherichia coli K-12 (bacteria)

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