Access to the cryoEM core.
Before you start working with our core be sure to complete the following check list:
- Have a iLAB account with accurate billing information. We use iLAB to book equipment and bill customers for services provided. UNC-CH researchers pay using Chartfeild string listed under there iLAB account. External customers can use credit card or a PO.
- Have Globus account setup. We use Globus to transfer data collected at our facility. CryoEM data is stored on the Dogwood/Longleaf HPC for 90 days (about 3 months). Data can be transferred from the Dogwood/Longleaf HPC using Globus.
- View our website to familiarize yourself with our policies and workflows.
- Setup meeting with the core director if you are interested in using our core facility.
- Have a cryoEM specific DMP for your lab or project. We will not store your data past 90 days. Most labs will keep a copy of the raw data on NSF or hard-drive and make duplicate copies.
Training and Learning CryoEM.
We offer training in all aspects of cryoEM to include specimen preparation of cryo-grids, operation of the cryoTEM, and image processing and analysis. Be sure to schedule an in-person or virtual meetings with the core director to discuss an individual training plan. Wednesdays are the preferred day for meetings between 11AM-6PM EST. We do not believe in “one-size fits all” for cryoEM and are happy to work with you on this. If you need help getting started, please refer to our website for resources and protocols.
Only staff that are currently employed by UNC-CH CryoEM Core can officially train a person to operate equipment at our core facility. We encourage individuals to help each-other, share knowledge openly with one another and learn from each other when they are working at the core. This includes watching a person operate the Vitrobot and microscope. This is most useful for labs that have multiple cryoEM projects. However, if you want to be able to operate the equipment at our core you must be fully trained by a core staff and have the permission of the core director.
Cryospecimen preparation using the Vitrobot.
Other than CryoEM Core Staff, the only individuals that have been fully trained by core staff can operate the Vitrobot. Vitrobot is available during normal Core hours M-F 9AM-7PM. Individuals that have been fully trained to operate the Vitrobot can use it after hours or over the weekend. The Vitrobot can be booked on iLAB using the Schedule Equipment tab. Customers are expected to (1) log the number of hours used in the Vitrobot notebook, (2) clean up after they complete and experiment, and (3) tell the core staff if they notice an error or problem with the equipment. The core can provide individual labs with assistance in preparing TEM support films including (1) graphene or graphene oxide, (2) thin carbon, or (3) lipid monolayers but are expected to be listed as collaborators if they help. The core can provide training for individuals interested in making their own TEM grid support films as a fee-for service.
We expect customers to have their own TEM grids if they are using the cryoEM Core. The core has grids in stock that can be given to customers or used to complete an experiment but only at the decision of the core director.
Long-term Storage of Cryo-grids at Our Core.
Customers that are using the cryoEM core to prepare or image grids can store their cryo-grids on site. Customers pay an annual fee based on the number of pucks they have at our core. Customers can request multiple pucks. We recommend using one puck per person, and one cane per lab. Customers are allowed to use grids boxes we keep at our core but are expected to return them once they have completed their experiment. Individual labs that have purchased their own cane and pucks are expected to use their own grid boxes.
CryoEM Core Staff and only individuals that have been fully trained by Core Staff can operate the Talos Artica. Individuals interested in using the Talos Artica will meet with the core director and develop an individual training plan for them to follow. We understand each person is unique with respect to their knowledge, project, goals, and experience. However, we expect that each person will respect the core staff, policies, and be held to the same high standards. Core Staff can operate the microscope for individuals or can complete an experiment for a customer as a fee-for service. Customers that rely on the CryoEM Core Staff for training and for operating the cryoTEM to complete an experiment will be billed accordingly. For routine research we bill Core Staff time as “Labor Fee”, anything beyond routine is consider a “Core Director Fee”. Core Staff can operate the microscope for individuals that are not able, willing, or interested in learning how to use it, as well as training for customers. If a core staff operates the Talos Arctica for a customer, they will be billed based on the number of hours the microscope is used in addition to the time the staff person spends operating the microscope. The total cost of the session will be microscope beam time + labor, additional fees or add-on charges would apply.
Trained users can operate the microscope on their own, however, some activities are strictly the responsibility of the core staff. These include loading the cassette into the microscope, advance microscope alignments, troubleshooting errors, contacting TFS or Gatan to service or maintain the microscope, and installing or setting up software. Customers are welcome to watch and learn, we will teach anyone if they are interested, but they are not permitted to do these activities.
Cassette exchanges are done by core staff and not customers. The core will exchange the cassette up to 2 times each day, with a third exchange possible with the permission of the core director. Normally we exchange the cassette before the morning session starts at 9AM and a second time at 2PM for the evening. More than two cassette exchanges per day are possible, however this is an inconvenience for us, and the customer will be billed for staff time.
When using the microscope be sure to take good notes and record your session information in the microscope log notebook. We use this notebook for billing. If you are unwilling or unable to do this a staff person will do so for you and you will be billed accordingly.
Booking and Scheduling the CryoTEM.
We use a queue for booking the cryoTEM. Individuals submit cryoTEM requests on iLAB which are reviewed by the cryoEM core each week. We expect the information in the request to be accurate, and it is the responsibility of the customer to complete the request. We are happy to help or fill out a request for the customer if requested by the customer.
Each week the microscope scheduled is prepared by the core director and staff. Sessions are booked based on order the request were received, priority of request, and availability of staff to help if required. Researcher submitting the request can specify a preferred day and time slot. We do not normally book the microscope more than 2 weeks in advance. Customers can cancel their session after contacting the core director, we prefer to know 1 day in advance however we understand things happen. If the person does not show up or cancels the day of the session, we consider this an inconvenience and will charge a late fee. If a customer cancels their session, we consider their request completed, and to book another session that person would need to submit a new request.
Cancelations in the event of a microscope error.
In the event of a critical microscope error any sessions that are canceled will be re-scheduled before booking other customers. If the customer does not want to be re-scheduled or prefers a later day, we will book other people that have submitted a request. The core will postpone making the schedule until the microscope is fully operational.
Customers can use the microscope in blocks of time: 9AM-2PM, 2PM-7PM, 2PM-9AM, or all day. If the customer requests 2PM-9AM or all-day session and they do not use the full 19- or 24-hour time slot they will be billed only for the beamtime used. However, we do charge a beam time minimum. If the customer books the microscope for 2PM-9AM session (19 hours) and uses less than 5 hours, they will be billed for 5 hours (corresponding to the 2PM-7PM time slot). If the customer books the microscope all day 9AM-9AM and uses the microscope for less than 10 hours, they will be billed for 10 hours (corresponding to 9AM-7PM). Customers that use less than 5 hours during the 9AM-2PM session are billed based on the number of hours they use the microscope.
We expect the grids listed on the cryoTEM request to be accurate. We understand mistakes happen or the project goals of the experiment change. We do our best to accommodate all our customers by being flexible. However, we consider loading cryo-grids not listed in the cryoTEM request as an inconvenience to the Core Staff. As such we will charge the customer an inconvenience fee. Customers can modify the request on iLAB to verify that the information is accurate, and the correct grids are listed.