Electron cryo-microscopy of biological specimens on conductive titanium-silicon metal glass films.
Thin films of the metal glass Ti88Si12 were produced by evaporation and characterized by AFM and conductivity measurements. Thin Ti88Si12 support films for electron microscopy were prepared by coating standard EM grids with evaporated films floated off mica, and characterized by electron imaging and electron diffraction. At room temperature, the specific resistance of a thin TiSi film was 10(6) times lower than that of an amorphous carbon film. At 77K, the specific resistance of TiSi films decreased, whereas that of carbon became immeasurably high. The effective scattering cross-section of TiSi and amorphous carbon for 120 kV electrons is roughly equal, but TiSi films for routine use can be approximately 10 times thinner due to their high mechanical strength, so that they would contribute less background noise to the image. Electron diffraction of purple membrane on a TiSi substrate confirmed that the support film was amorphous, and indicated that the high-resolution order of the biological sample was preserved. Electron micrographs of TiSi films tilted by 45 degrees relative to the electron beam recorded at approximately 4 K indicated that the incidence of beam-induced movements was reduced by 50% compared to amorphous carbon film under the same conditions. The success rate of recording high-resolution images of purple membranes on TiSi films was close to 100%. We conclude that TiSi support films are ideal for high-resolution electron cryo-microscopy (cryo-EM) of biological specimens, as they reduce beam-induced movement significantly, due to their high electrical conductivity at low temperature and their favorable mechanical properties.
Rhinow
Daniel
D
Max Planck Institute of Biophysics, Max-von-Laue-Strasse 3, D-60539 Frankfurt am Main, Germany.
Kühlbrandt
Werner
W
D020285:Cryoelectron Microscopy; D005898:Glass; D018717:Purple Membrane / Q000648:ultrastructure*; D012825:Silicon; D013814:Thermal Conductivity; D016707:Tissue Fixation / Q000379:methods*; D014025:Titanium
D014025:Titanium; D012825:Silicon