Equipment & Methodology

In our space located on the fifth and second floors of the Black building, we have all the equipment to prepare our samples, collect the EM data and analyze them:

Grid preparation:

  • Edwards Auto 306 Turbo-pumped carbon evaporation system. This tool creates a clean, hydrophilic and non-reactive surface which is needed for making frozen-hydrated grids of labile macro-molecular complexes
  • Manual two-sided cryo plunging unit. This system creates frozen-hydrated samples reproducibly with minimal post-blotting cooling.
  • FEI Vitrobot Mark IV Vitrification robot. (Arrival December 2008). With this cryo-plunging system, parameters such as blotting time, relative humidity and temperature can be controlled for reproducible results even by inexperienced users.

Electron Microscopes:

  • FEI Tecnai F20 Cryo TEM with Gatan 4x4k ultrascan CCD camera and CT3500 cryo-transfer system (Arrival 1/2009). This microscope operates with voltages up to 200 kV and has a highly coherent electron source for high-resolution data collection from frozen-hydrated samples. A number of software packages have been installed to allow automated data collection.
  • FEI Tecnai Polara F30 Cryo TEM with Tietz 4x4k CCD camera (Arrival 10/2008). This microscope operates with voltages up to 300 kV and is capable of operating at liquid helium or liquid nitrogen temperatures. The instrument has a very stable cartridge style system that allows up to 6 samples to be loaded at once for screening. Once the best sample has been found, data can be collected for multiple days with little contamination or operator intervention using a program developed in house, called AutoEMation (see Lei, J., and Frank, J. (2005) in publication list).
    Vibration and field canceling systems are also installed. Click on the image below for an illustration of their effects:

    New: F30 calibrations by SerialEM.

Data Digitization and Screening:

  • SIRA Digital diffractometer. Data can be quickly screened for image quality using this apparatus.
  • ZI Photoscan 2001. This high-resolution (7 micron) scanner allows data collect on film to be digitized with great accuracy for high-resolution results.

Data Processing:

  • SPIDER image processing system,
  • Cluster and computing facilities,