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Electron Tomography for cellular organelles.
 

Options:

  1. Frozen cells cultured on the holey carbon grid
  2. Plastic cellular sections embedded in the epoxy resin
  3. Platinum replicas for the membrane undercoats
     

Workflow:

  1. Wash holey carbon membrane/ gold grids with pure acetone (overnight at least)
  2. Natural dry
  3. Glow discharge the grid by the Quorum GloQube
  4. Culture the tissue cells on the gold grid
  5. Blot the access volume on the grid with the single filter paper
  6. Vitrification purified protein-complexes on the grid by the FEI Vitrobot
  7. Set the frozen grid on the Gatan cryo-transfer holder
  8. Insert the cryo-holder to the FEI TalosF200C
  9. General alignments for TEM
  10. Adjust auto Eucentric height & auto focus at optimised position
  11. Calibrate the Gatan or Fischione sample holder
  12. Calibrate the dynamic focus (for STEM-tomography)
  13. Prepare the acquisition positions (for cryo-tomography)
  14. Prepare step conditions before starting a tilt series
  15. Acquire tilt series
  16. Reconstruct live tilt series
  17. 3D-reconstruction and analysis by Inspect3D or IMOD

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