Vanderbilt Cryo Electron Microscopy Facility
The V-CEM facility provides expertise and resources for the structural analysis of biological molecules using single particle analysis, tomography, and two-dimensional (2D) crystallization. The facility houses high-resolution transmission electron microscopes (TEMs) and cameras needed to collect data for single particle analysis and tomography.
The facility houses a 300 kV FEI Polara equipped with a Gatan K2 Summit Direct Electron Detector (DED) camera, a 200 kV FEI F20 equipped with a Gatan Ultrascan 4K x 4K CCD, two FEI vitrobots (mark II\III), and the accompanying instrumentation needed for both negative stain and cryo-EM analysis. Biological samples are prepared for molecular EM using either negative stain or vitrification (i.e., cryo-EM). For negative stain EM, the sample is embedded in a layer of dried heavy metal solution. For cryo-EM, the sample is suspended across a holey carbon grid and then rapidly plunged into cryogen, trapping the molecules in a thin layer of vitrified ice composed of the purification buffer. Negative stain provides high-contrast structural information mainly of the outer envelope of a molecule, while cryo-EM allows for the structural determination of near-atomic to atomic resolution 3D structures.
The goal of V-CEM is to make TEM accessible to all interested researchers at Vanderbilt. TEM is one of the fastest developing, cutting-edge structural techniques for exploring the organization of biological macromolecules. The unique strength of TEM is its versatility. Structures determined using TEM can span resolution ranges from very low- to atomic-levels (>30–1.8Å) and do not require crystalline samples for analysis.
In pursuit of its goal, the V-CEM enables TEM training and outreach by:
- Providing extensive training to new users.
- Providing advise and support including specimen preparation, data collection and image processing, computational image analysis, and structure determination.
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