Any use of the resources at the Fred Hutchinson Cancer Research Center Cryo-EM shared resource requires acknowledgement of the core in publication as follows: “Electron microscopy data were generated using the Fred Hutchinson Cancer Research Center Cryo-EM shared resource.” Descriptions of the overall Fred Hutch Shared Resources program are available on the main Shared Resources grant information page.
The Cryo-EM shared resource provides labs with training and access to two high-resolution transmission electron microscopes and the necessary sample-preparation tools. This shared resource offers negative-stain and vitrification tools, a Talos L120C TEM, a Glacios 200kV TEM, four A6000 GPUs, automated data-collection software, and staff trained in high-resolution structure determination by cryo-EM.
The Cryo-EM shared resource provides users with the support and resources needed to determine structures of biological specimens to further their scientific pursuits. The electron microscopy suite at Fred Hutchinson Cancer Research Center is newly renovated to accommodate 200 kV Glacios and 120 kV Talos transmission microscopes. These are available for assisted and independent use after training provided by staff members in the Cryo-EM shared resource. In the same approximately 550 square feet of space, all tools and instruments needed for negative stain and sample vitrification on the Mark IV Vitrobot are provided for labs to use. Shared resource staff maintain the microscopes and sample-preparation equipment in a fully functional state with alignments verified bimonthly. They also train new and experienced users in the operation of the hardware and software provided by the core, detailed below.
The Talos microscope is equipped with a Ceta 16M CMOS camera for high-speed and high-resolution 4k x 4k imaging in low-dose mode. This microscope is used for negative-stain data collection and cryo-EM screening with our room-temperature and cryogenic Elsa holder. The Glacios microscope is equipped with a K3 camera, the newest generation of direct electron detector from Gatan. This camera has a large sensor that can operate at 1500 frames per second, allowing for exceptionally sensitive and fast data collection.
The shared resource is also supported by Fred Hutch Shared Resources and Center IT to provide 10 GbE data-transfer connections, enabling on-the-fly drift correction and initial live processing during imaging sessions on the Glacios. All of these data are retained for one month on the shared resource’s storage server, and users are provided with options for transferring data off this server internally and externally. Both microscopes have SerialEM and Leginon installed and calibrated for use in automated data collection, which enable the collection of dozens of micrographs per hour. Image processing is supported by individual labs, but staff provide office hours and individual consultations on adjusting data-processing parameters for optimal results.