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FEI Tecnai F30 G2 Twin Microscope

FEI Tecnai F30 Twin is a 300kV field emission gun (FEG) high resolution TEM/STEM which was installed in May 2012. The microscope has a Twin objective lens with a Cs of 2.0 mm. With its 300 kV acceleration voltage and its Schottky field emission source, this instrument provides outstanding stability, ease-of-use and high-resolution performance in TEM/STEM imaging and microanalysis.
The scope is equipped with an Oxford ultra-thin window EDX detector, a Gatan ultra-scan 4000 CCD and Fischione high angle annular dark field detectors for STEM imaging, Quantum Gatan Imaging Filter with 2Kx2K CCD for performing EELS and energy filtered imaging, Fischione tomography holder, Xplore3D Xpress tomography data collection software (batch tomography), ResolveRT batch processing tomography software and AMIRA 3D rendering and segmentation software, Gatan 626 cryo-holder with α +/- 70 º tilt range and ability to keep samples at -170 ºC for ~4 hours, Gatan pumping station, Gatan cryo-blades, low-dose data collection package, Mark IV Vitrobot for making vitreous plunge-frozen samples.

  • Operating voltages of 100, 200 and 300 kV
  • Point resolution: 0.24 nm
  • Information limit: 0.14 nm
  • Energy resolution in EDX: 134 eV
  • Magnification range: 58x - 800 Kx in TEM mode; 150-230 Kx in STEM Mode
  • Magnification reproducible within ± 1.5%
  • Camera Length range: 100 – 5600 mm in TEM-diffraction mode
  • Coma-free alignment for high-resolution objective-lens centering
  • Rotation-free magnification and diffraction series
  • Fully computer-controlled, eucentric side-entry, high stability CompuStage
  • Double tilt low background analytical specimen holder (+/- 70º)
  • Excellent EDS in-hole performance, low system background in EDX, small probes and embedded EDX, spectrum profiling and imaging
  • Improved EDS analytical capabilities through specimen drift correction control
  • Xplore3D Xpress Tomography with high tilt of α +/- 80 º
  • Quantum GIF for performing EELS/EFTEM
  • Cryo-EM with Gatan 626 high tilt Cryo-holder

 

Cryo-electron microscopy

  • Rapid freezing results in thin vitreous(glass-like) ice.
  • Frozen, hydrated native state
  • Visualization of proteins (>100 kDa) in solution,
  • 5-50 Angstrom structure determination

(Left from White, 2010 PLoS) 2D Projection Images of SIV in vitreous ice.  (Middle) The B. stearothermophilus pyruvate dehydrogenase E2 core domain.  (Right) Model of pyruvate dehydrogenase complex.

XEDS
(X-ray Energy Dispersive Spectroscopy)

  • Detect X-rays generated and identify elements (sodium-germanium)
  • Useful in determining which elements are present


(Left from Holman, 2008 PNAS) Cryo-EM of Desulfovibrio containing electron dense material. (Middle) Elemental composition of electron dense using XEDS.  (Right) Experimental display of locations of various elements during XEDS.


Tomography

  • Acquisition of tilt series +/- 70o, dual axis
  • From tilt series of 2-D projection images can generate 3D volume
  • Useful in determining localization, orientation, shape & distribution

(Left from White, 2010 PLoS) Tomographic reconstruction of SIV virion. (Middle) Segmentation of tomographic reconstruction. (Right) 3D classification and averaging of 3D SIV Envelope glycoprotein subvolumes

Electron Diffraction

  • Collect diffraction data of crystalline specimens
  • 0.14 lattice and 0.24 point resolution
  • Useful in detecting material defects, characterizing nanocrystals, deterimining membrane protein structure

(Left from T. Gonen, 2004 JMB) Crystals of Aquaporin-0 and electron diffraction patterns of untilted double-layered AQP0 2D crystals typically showed diffraction spots to a resolution better than 3.5 Angstrom (Middle). (Right) Model of Aquaporin.

STEM 
(Scanning Transmission Electron Microscopy)

  • Finely probe nanometer diameters with electron beam
  • Elements with high atomic number (Z) display more contrast
  • High angle annular dark field (HAADF) and HR-STEM
  • Useful in detecting material defects, heavy atoms and gold labels

(Left from A. Herzing 2008 Science) High-magnification aberration-corrected STEM-HAADF images of (A and B) the inactive and (C and D) the active Au/FeOx catalysts acquired with STEM. (Right from Sousa, 2011 JSB) Axial STEM tomogram showing a 1-μm-thick section of pancreatic beta cells.

EELS
(Electron Energy Loss Spectroscopy)

  • Analysis of inelastic electron scattering
  • Shape of energies and ionization edges indicate elemental composition, chemical binding, valence, and conduction, electron density and sample thickness.
  • Elements with low atomic number (Z) are easier to detect