Located in the Morrison Center, the CryoEM Core Facility has a state-of-the-art Glacios 200kV electron microscope available for structure determination of protein, nucleic acids, and other molecules of biological interest, including small molecules in a crystalline state. CryoEM Core services include:
- Single-particle CryoEM;
- Cryo-electron tomography (CryoET) of thin specimens;
- Micro-crystal electron diffraction (MicroED);
- Sample preparation;
- Comprehensive data collection and analysis of CryoEM samples;
- Sample storage and shipping; and
- Data processing.
Instruments include:
- A 200kV Glacios CryoTEM: The Thermo Scientific Glacios cryo-transmission electron microscope (Cryo-TEM) allows users to easily collect near atomic data from a broad range of biological targets. Compared to the previous generation, the Glacios cryo-TEM delivers higher throughput and makes cryo-EM more accessible and efficient, thanks to the Aberration Free Imaging System (AFIS) and Fringe-Free Imaging ( FFI) system. The Glacios features a Falcon 4i direct electron detector, a Selectris energy filter, and the Thermo Scientific EPU software. In addition, the instrument can load up to 12 grids, reducing sample contamination and speeding up the screening process. Furthermore, the Glacios has the MicroED package, which includes a CetaD camera, optimized apertures, a smaller beam stop, and EPU-D. The Glacios cryo-TEM is ideally suited for single-particle analysis, cryo-electron (CryoET) tomography, and MicroED.
- A TFS Vitrobot Mark IV: The Vitrobot is a fully computer-controlled double-sided bloating robot for the vitrification of aqueous samples. Minimal training is required for operating the instrument in a laboratory environment. The process of plunging, blotting, and vitrification is fully automated upon placing a vial in the chamber and setting up the liquid coolant container. Operational parameters tend to be reproducible, enabling a high throughput of vitrified samples with a straightforward control of the vitrification process. The Vitrobot allows users to control the temperature, humidity, number of blotting, and several critical time settings. The instrument consistently produces excellent specimens for cryo-electron microscopy, including single-particle, CryoET, and MicroED.
- A Leica EM GP2: The Leica EM GP2 is a fully computer-controlled single-sided bloating vitrification robot. Bloating on the EM GP2 can be performed using sensor bloating or manual blotting using the onboard eyepiece. This instrument allows the user to perform multiple bloating operations, including back and front bloat. The possibility of back bloating makes the EM GP2 a perfect tool for the vitrification of cells and microcrystals. In contrast with the Vitrobot, the EM GP2 is capable of controlling the temperature of liquid ethane and providing a continuum flow of liquid nitrogen in the sample vitrification area, considerably reducing ice contamination. The instrument is highly reproducible and excellent for the three main CryoEM workflows, including single-particle CryoEM, CryoET, and MicroED.
- A PELCO easiGlow™: The PELCO easiGlow™ glow discharge cleaning system is a compact, quick, easy-to-use standalone system. It is designed for cleaning and hydrophilizing TEM carbon support films or grids, which tend to be hydrophobic. A glow discharge treatment with air makes a carbon film surface negatively charged (hydrophilic), which allows aqueous solutions to spread evenly. Glow discharge treatment of TEM grids removes adsorbed hydrocarbons, cleaning them while making them hydrophilic. The PELCO easiGlow™ supports hydrophilic and hydrophobic treatment with either negative or positive charge and includes two separately controlled gas inlets. The vacuum level in the instrument is precision-controlled electronically by a proportional valve, a novel approach that eliminates manual settings with a needle valve. A gentle venting procedure ensures that the TEM grids are not disturbed when the system is vented.
