Cryoelectron Tomography (CryoET) Centers
The National Network for CryoET was established as part of the Transformative High Resolution Cryo-Electron Microscopy Program in 2020, funding four centers to provide the biomedical research community access to advanced instrumentation for cryoET, cryoET specimen preparation, and collection of high-resolution cryoET data as well as cross-training in cryoET methods.
The network comprises a HUB, The Midwest Center for Cryo-Electron Tomography (MCCET) that performs high-resolution data collection and three “spokes”, or centers that provide optimal sample preparation and screening. The spokes are located at CU Boulder Center for Cryo-ET (CCET), The National Center for In-situ Tomographic Ultramicroscopy (NCITU), and Stanford-SLAC CryoET Specimen Preparation Service Center (SCSC). All of the centers are accessible through competitive peer review of user proposals administered by the HUB and there are no geographical restrictions.
Learn more and apply for access at www.cryoetportal.org
Network Hub: The Midwest Center for Cryo-Electron Tomography (MCCET)
Located at the University of Wisconsin-Madison, MCCET provides access and training in all aspects of the CryoET pipeline and is responsible for the coordination of the national CryoET network activities between the NIH, network centers, and users.
Types of access:
- Routine and advanced cryo-electron tomography (cryo-ET) specimen preparation, data collection, and computation
- Hands-on, remote, and virtual training in cryo-ET specimen preparation, data collection, and data processing and validation
- Academic and commercial data collection services
Instrumentation Current and Planned:
- 300kV Titan Krios G3i Cryo-TEM with Volta phase plates, STEM detector, Falcon 3C direct electron detector, Gatan Bioquantum Post-column Energy Filter with K3 direct electron detector, Ceta CMOS camera.
- 300kV Titan Krios G4i Cryo-TEM with Selectris Energy Filter and Falcon 4i direct electron detector.
- 200kV Talos Arctica with Volta phase plates, STEM detector, Falcon 3C direct electron detector, Gatan Bioquantum Post-column Energy Filter with K3 direct electron detector, Ceta CMOS camera.
- 120kV Talos L120C with Ceta CMOS camera, room temperature specimen holders, Gatan 626 cryotransfer system and holders.
- Aquilos 2 Cryo-FIB-SEM (MCCET) with dedicated cryo-stage cooling, a loading and transfer system, field emission SEM column. Integrated fluorescence imaging (iFLM) for correlative light microscopy.
- Aquilos 2 Cryo-FIB-SEM (CEMRC) with dedicated cryo-stage cooling, a loading and transfer system, field emission SEM column without iFLM.
Fluorescent Light Microscopes:
- Leica DMi8 widefield fluorescence microscope with Alveole PRIMO Micropatterning system.
- Leica M165 Stereo microscope.
- Leica Cryo-CLEM System with cryo-CLEM stage and transfer shuttle, Leica DM6 FS automated wide-field upright microscope and motorized stage; filter systems for DAPI, GFP, Texas Red (mCherry), YFP, CFP, Cy3, and Cy5. Software for automation and image processing.
- Leica Stellaris Five Cryo Confocal microscope.
Sample Preparation:
- FEI Vitrobot Mark IV cryo-plunge freezing system.
- Leica EM GP2 Cryo-plunge freezing system.
- Leica ACE600 System for glow discharge, carbon evaporation, and metal coating.
- Leica EM ICE High pressure freezing system.
- Leica Ultracut Cryo-Ultramicrotome.
- Cryo-grid storage with puck holders
- Dedicated wet and dry lab research spaces for sample handling.
- Stereo microscopes
- Shaking incubators, CO2 incubators
- Biosafety cabinets
CU Boulder Center for Cryo-ET (CCET)
Located at the University of Colorado, CCET will focus on cryo-specimen preparation and cryo-electron tomography (cryo-ET) applied to cells, organelles and large supramolecular assemblies.
Types of access:
- HPF sample preparation
- VitroJet sample preparation
- FIB-SEM sample preparation
- THUNDER-CLEM sample correlation
- Time resolved sample preparation
- Sample screening
- Embedded User Training
Instrumentation Current and Planned:
- Aquilos-2 CryoFIB
- Thunder CLEMVitrobot Mark V
- Cryosol Vitrojet
- Wohlwend High Pressure Freezer
- Tecnai F30; K2; OneView
- Tecnai F20; K3
- GPU work stations
The National Center for In-situ Tomographic Ultramicroscopy (NCITU)
Located at the New York Structural Biology Center, NCITU will be dedicated to developing and providing access to both standard and advanced methods for in-situ cryoET specimen preparation, and to disseminating this expertise through a cryoET cross-training program designed to serve a wide variety of skill levels and career goals.
Stanford-SLAC CryoET Specimen Preparation Service Center (SCSC)
Located at Stanford University, SCSC will enable external users to streamline the preparation of samples through providing access to, and support for, advanced cryo-specimen preparation techniques (cryoFLM, cryoFIB, and cryoEM/ET) for a wide range of samples with biomedical applications. The Center will offer a comprehensive program to train and cross-train scientists, regardless of their institutional affiliations, to become independent cryoET investigators.