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Center for Structural Biology

Equipment

CryoTEM ThermoFisher Scientific Talos Arctica

  • 200 keV transmission electron microscope.
  • FEG-X Gun Schottkey Emitter.
  • Two Condenser Lens system.
  • Constant power C-Twin object lens with Cs = 2.7 mm.
  • Autoloader capable of holding 12 cartridges.
  • Ceta 4096 x 4096 pixel CMSO CCD.
  • Gatan K3 5,760 x 4,092 pixel direct electron detector.
  • Automated SP and cryoET data collection using SerialEM.
  • On-the-fly processing using cryoSPARC Live and Warp
  • Capable of obtaining near atomic resolution for single-particle cryoEM.
  • Benchmarked with Apoferritin sample 1.8 Å resolution based on FSC=1.34.

If you would like to learn how to operate the Talos Arctica
  1. Setup iLAB account with accurate billing information. 
  2. Read CryoEM101 — Chapter 3 Grid Screening and Evaluation, Chapter 4 Data collection
  3. Watch Grant Jensen CryoEM Course – Unit 1 TEM
  4. Read UNC CryoEM Talos Arctic Protocol
  5. Contact the Core Director and ask to schedule a training session. 
  6. Once the Core Director feels you are competent to operate the microscope you will be able to use it without supervision and after hours.

Vitrobot Mark IV

  • Used to prepare cryo-grids by rapid immersion in liquid ethane/propane mixture.
  • Located in Glaxo Research Building room 011 across the hall from the Talos Arctica.
  • Available for general use M-F 9 AM-7 PM.  If you would like to use the Vitrobot after-hours or over the weekend, please contact the Core Director.

Pelco easiGlow

  • Used to render TEM grids hydrophilic by glow discharging (plasma cleaning with air).
  • Located in the cryo-prep room across the hall from the microscope control room.
  • Custom presettings optimized for Quantifoil TEM grids.

Tergeo-EM plasma cleaner

  • Plasma cleaner is capable of producing indirect or direct plasma derived from argon and oxygen.  Used to render UltrAufoil and Quantifiol TEM grids hydrophilic.
  • Custom recipes for CryoEM applications for different types of TEM grids including (1) Quantifoil, (2) Lacey Carbon, (3) TEM grids with continuous layer of thin 2-nm carbon, (4) C-Flat, (5) UltrAufoil TEM grids.

Cryogrid Storage
ThermoFisher Biocane 34 and Subangstrom Pucks
  • Two Liquid nitrogen dewars used for long-term storage of cryo-grids.
  • Maintained by the cryoEM Core Staff.
  • Can accommodate 6 cane (60 pucks: 600 cryogrid boxes: 2,400 cryogrids).
  • If you like to store your samples at the core, please let us know.

Leica EM ACE600 Sputter Coater

  • High vacuum coating system used to prepare (1) thin amorphous carbon films and (2) UltrAufoil style gold films for cryoEM applications.  
  • Located in Glaxo Room 036 near the Vitrobot.

Clipping Station

  • ThermoFisher Scientific clipping station located in the microscope control room.
  • Everything you need to clip TEM grids including c-clips and rings. 
  • Clipping tools, forceps, liquid nitrogen dewars, and drying station, and forceps.

 

If you would like to learn how to prepare your own cryo-grids:
  1. Setup iLAB account with accurate billing information. 
  2. Read CryoEM101 — Chapter 1 Sample Preparation and 2 CryoEM Grid Preparation. 
  3. Watch Grant Jensen CryoEM Course – Unit 2 Sample Preparation. 
  4. Read our CryoEM Vitrobot Protocol. 
  5. Contact the Core Director and ask to schedule a training session. 
  6. Once the Core Director feels you are competent to operate the Vitrobot you will be able to use it without supervision and after hours.