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TitleReducing effects of particle adsorption to the air-water interface in cryo-EM.
Journal, issue, pagesNat Methods, Vol. 15, Issue 10, Page 793-795, Year 2018
Publish dateSep 24, 2018
AuthorsAlex J Noble / Hui Wei / Venkata P Dandey / Zhening Zhang / Yong Zi Tan / Clinton S Potter / Bridget Carragher /
PubMed AbstractMost protein particles prepared in vitreous ice for single-particle cryo-electron microscopy (cryo-EM) are adsorbed to air-water or substrate-water interfaces, which can cause the particles to adopt ...Most protein particles prepared in vitreous ice for single-particle cryo-electron microscopy (cryo-EM) are adsorbed to air-water or substrate-water interfaces, which can cause the particles to adopt preferred orientations. By using a rapid plunge-freezing robot and nanowire grids, we were able to reduce some of the deleterious effects of the air-water interface by decreasing the dwell time of particles in thin liquid films. We demonstrated this by using single-particle cryo-EM and cryo-electron tomography (cryo-ET) to examine hemagglutinin, insulin receptor complex, and apoferritin.
External linksNat Methods / PubMed:30250056 / PubMed Central
MethodsEM (tomography) / EM (single particle)
Resolution3.77 - 6.09 Å
Structure data

EMDB-7623:
Apoferritin with spot-to-plunge time of 100ms
Method: EM (tomography)

EMDB-7624:
Hemagglutinin with spot-to-plunge time of 800ms
Method: EM (tomography)

EMDB-7625:
Apoferritin with spot-to-plunge time of 400ms
Method: EM (tomography)

EMDB-7627:
Apoferritin 0.5 mM TCEP with spot-to-plunge time of 170ms
Method: EM (tomography)

EMDB-7628:
Hemagglutinin with spot-to-plunge time of 100ms
Method: EM (tomography)

EMDB-7629:
Insulin-bound insulin receptor with spot-to-plunge time of 200ms
Method: EM (tomography)

EMDB-7630:
Insulin-bound insulin receptor with spot-to-plunge time of 600ms
Method: EM (tomography)

EMDB-7788:
Insulin Receptor ectodomain in complex with two insulin molecules plunged with a spot-to-plunge time of 200 ms
Method: EM (single particle) / Resolution: 4.93 Å

EMDB-7791:
Insulin Receptor ectodomain in complex with two insulin molecules plunged with a spot-to-plunge time of 800 ms
Method: EM (single particle) / Resolution: 6.09 Å

EMDB-7792:
Influenza hemagglutinin trimer plunged with a spot-to-plunge time of 100 ms
Method: EM (single particle) / Resolution: 3.77 Å

Source
  • Tabanus atratus (black horsefly)
  • unidentified (others)
  • Homo sapiens (human)
  • Influenza A virus (A/Hong Kong/26/1983(H3N2))

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