Training
Training for cryoEM at The Hormel Institute is based on the National Institutes of Health Common Fund CryoEM Initiative Merit Badge curriculum. We strongly recommend you review the NIH’s CryoEM/CryoET 101 tutorials to be prepared for essential knowledge you will need to showcase your capabilities before using cryoEM equipment at The Hormel Institute.
To earn a merit badge, you will need to:
- Conduct independent study by reviewing background information and SOPs
- Schedule a practical demonstration with The Hormel Institute cryoEM staff
- Schedule a hands-on practice session with The Hormel Institute cryoEM staff
- Complete an in-person practical test
Once you have received your merit badge, you will be able to use cryoEM equipment and shared resources at The Hormel Institute.
While merit badges earned at other facilities will be honored, users will still need to demonstrate competency with the equipment by completing an in-person practical test at The Hormel Institute.
Essential Base Knowledge and SOP
The Vitrobot’s primary function is to create thin, vitreous layers of biological samples, such as proteins or cells, suitable for cryoEM imaging. The process involves applying a small amount of the sample onto a grid, removing excess liquid through blotting, and then rapidly plunging the grid into a cryogen (typically liquid ethane). The Vitrobot automates the blotting step, which is crucial for controlling ice thickness and removing excess liquid from the grid. Precise control over parameters ensures that the vitrification process is highly reproducible, resulting in more reliable and consistent cryoEM data.
Knowledge Test
Follow this link https://www.cryoemcenters.org/merit-badge/
Videos and Information
Essential Base Knowledge and SOP
Grid clipping prepares cryo-EM samples for automated loading and imaging in a cryo-TEM equipped with an autoloader. A cryo-EM grid is placed on a specialized metal ring and secured with a C-shaped clip, forming an autogrid. This assembly is done at a dedicated clipping station, often cooled, to keep grids and tools cold and prevent ice contamination. Clipped grids are then loaded into autoloader cassettes, which enable fully automated, high-throughput sample exchange inside the microscope.
Knowledge Test
Follow this link https://www.cryoemcenters.org/merit-badge/
The pass/fail knowledge assessment will test understanding of the following concepts:
- Why good clipping is important for your grids and the microscope
- The purpose of PPE for grid clipping
- Keeping your grids vitrified during clipping
- Determining if your grid is clipped properly
- ID tools used for grid clipping
Here is the current quiz version:
Videos and Information
Log Sheets and Checklists
Essential Base Knowledge and SOP
CryoEM autoloader systems simplify and automate the process of introducing samples into cryoelectron microscopes, minimizing manual handling and lowering the risk of sample contamination or damage. To prepare for this, cryo-EM grids are clipped and these clipped grids are placed into an autoloader cassette, which can hold multiple grids at once. Once the cassette is inserted into the microscope, the autoloader’s robotic mechanism transfers each grid individually from the cassette to the microscope’s specimen stage for imaging. After data collection, the system can automatically retrieve and store the grids back in the cassette, increasing efficiency and reducing the chance of handling errors.
Knowledge Test
Follow this link https://www.cryoemcenters.org/merit-badge/
Log Sheets and Checklists
Essential Base Knowledge and SOP
The Tecnai G2 Spirit BioTWIN is a type of Transmission Electron Microscope (TEM) used in biological and materials science research. It is known for its high-contrast and high-resolution imaging capabilities, making it suitable for examining a wide range of samples, including negatively stained protein samples, biological samples such as cells and tissues.
Videos and Information
Log Sheets and Checklists
Essential Base Knowledge and SOP
The Cryoplunge CP3 is a Gatan instrument used to prepare samples for cryo-EM. It is a semi-automated device that rapidly vitrifies protein or other biological samples for cryo-electron microscopy imaging. The system includes a humidity-controlled chamber and features timed blotting for precise sample preparation. The Cryoplunge CP3 provides consistent, high-quality, vitrified grids for cryo-EM imaging.
Log Sheets and Checklists
Essential Base Knowledge and SOP
EPU stands for Automated Data Collection Software from Thermo Fisher Scientific. It's designed to streamline and automate many tasks, such as selecting suitable areas on the cryo-EM grid for imaging, adjusting microscope settings, and monitoring imaging conditions, increasing efficiency and throughput. EPU is a key tool in the cryoEM workflow, particularly for single particle analysis, where it helps researchers collect large datasets of images of individual molecules.
Knowledge Test
Follow this link https://www.cryoemcenters.org/merit-badge/
Log Sheets and Checklists
Essential Base Knowledge and SOP
Single-particle cryo-electron microscopy (cryoEM) data processing is a powerful computational workflow used to determine high-resolution 3D structures of biological macromolecules. The process starts with motion correction and contrast transfer function (CTF) estimation of raw micrographs. Individual particle images are then picked and extracted for further analysis. 2D classification helps identify homogeneous particle views and remove contaminants or damaged particles. High-quality particles are subjected to 3D map generation. This workflow enables us to visualize protein complexes, viruses, and large assemblies at near-atomic resolution. The resulting maps can be interpreted to build accurate atomic models of the target molecule.
Data Processing Standard Operating Procedures