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Single Particle Analysis for purified protein complexes.
 

Workflow:

  1. Wash holey carbon membrane/ gold grids with pure acetone (as required)
  2. Natural dry
  3. Glow discharge the grid by the Quorum GloQube
  4. Prepare graphene-oxide membrane on the gold grid (as required)
  • Graphene oxide dispersion in H2O (Sigma-Aldrich, Merck)
  • Quantifoil, C-flat, UltraAuFoil
  • Anti-capillary crossover tweezers
  1. Prepare ultra-thin carbon membrane on the gold grid (as required)
  • Carbon coats on mica
  • Quantifoil, C-flat, UltraAuFoil
  1. Put the purified protein-complex on the designed grid as above
  2. Blot the access volume on the grid with filter papers
  3. Vitrification purified protein-complexes on the grid by the FEI Vitrobot
  4. Set the frozen grid on the Gatan cryo-transfer holder
  5. Insert the cryo-holder to the FEI TalosF200C
  6. General alignment for low electron-dose imaging
  7. EPU image acquisition after the Atlas
  8. Image processing by the Relion or CryoSparc on the Cryo-EM GPU server
  9. Repeat several refinements
  10. 3D-model

Group Members

Jaime Llodra Gonzalez

EM Research Specialist

jl2235@mrc-tox.cam.ac.uk

 

Maria Guerra Martin

Senior Research Officer

mag88@mrc-tox.cam.ac.uk

 

Nobuhiro Morone

Senior Investigator Scientist

Facility Head

nm669@mrc-tox.cam.ac.uk