We offer fee-for-services for (1) cryo-grid sample preparation using a Vitrobot Mark IV, (2) Screening and Data collection using Talos Arctica CryoTEM. \u00a0Booking equipment and billing are done in iLAB, make sure to have an account set up before using the CryoEM Core. \u00a0If you would like to learn how to operate the Vitrobot and Talos Arctica, please contact the Core Directory directly and request training.<\/p>\n
Pre-processing of single-particle cryoEM data, including motion correction and CTF estimation using Warp or CryoSPARC Live, is not considered “image processing” and is typically included in the data collection. \u00a0Anything beyond this is considered image processing, including 2D classification and EM map generation. \u00a0Having the ability to process data on the fly is incredibly useful, and we routinely will generate 2D class averages and 3D models to determine the structure of the target complex as it is being collected. \u00a0If you need assistance with this, we are happy to help, we only ask that you acknowledge the CryoEM Core Staff that process the data. \u00a0Image processing can be done during data collection and typically takes a few hours to complete.<\/p>\n
Only trained users are permitted to operate the Vitrobot. \u00a0The Vitrobot is located in Glaxo Research Building room 011 across the hall from the Talos Arctica. \u00a0The Vitrobot Mark VI can be reserved using iLAB. \u00a0If you need assistance or require training please contact us directly.<\/p>\n
The Talos Arctica is used for cryo-EM specimen screening and<\/span> high-resolution data collection. \u00a0Only trained users are permitted to operate the Talos Arctica. \u00a0Researchers interested in getting trained or using the microscope, please contact the\u00a0<\/span>cryo-EM<\/span>\u00a0core director Dr. Joshua Strauss. \u00a0Only\u00a0<\/span>cryoEM<\/span>\u00a0core staff\u00a0<\/span>are allowed to<\/span> dock\/undock the cassette into the Autoloader and load\/unload grids into the cassette. \u00a0We limit the number of docking\/<\/span>undockings<\/span> to two per day and only during working hours of the core 9AM-7PM. \u00a0\u00a0We typically schedule two sessions per day, M-F.\u00a0 The morning session, 9AM-2PM, is only for screening samples with the option to collect a small data set of up to 1000 movies (about 2 hours of data collection). \u00a0 The evening session, from 2PM-7PM, is for either (1) screening samples with the option to collect ~5,000 movies overnight, or (2) for collecting data overnight. \u00a0If you require a dataset larger than 5000-6000 micrographs, data can also be collected over the weekend.\u00a0 We will typically load the sample Friday afternoon and monitor data collection remotely.<\/span><\/span>\u00a0<\/span><\/p>\n Each core user is responsible for the long-term storage of their data.\u00a0 Data collected from the Gatan K3 camera is stored in the Dogwood\/Longleaf research computing cluster, where it can be accessed via Globus. \u00a0We do not offer long-term storage (> 3 months) for data collected on the Talos Arctica TEM. \u00a0Additional information about the Dogwood and Longleaf computer clusters are on the UNC Research Computing website https:\/\/its.unc.edu\/research-computing\/techdocs\/longleaf-frequently-asked-questions-faqs\/<\/a><\/p>\n Core users can store cryo-grids in the Biocane34 liquid nitrogen dewar located in the cryo-prep room at Glaxo Research building. \u00a0The cryoEM core maintains a record of the samples stored in this dewar, however, we encourage people to keep a duplicate record of the location of their samples. \u00a0Only cryoEM core staff are permitted to remove and place samples in the Biocane34 dewar. \u00a0Each lab is allowed one puck to store their cryo-grids. \u00a0If you need more than one puck, contact the Core Director.<\/p>\n <\/p>\n","protected":false},"excerpt":{"rendered":"Data management<\/h2>\n
Storage of Cryogrids<\/h2>\n