Policies/Instructions

Janelia CryoEM Facility Policies

The Janelia CryoEM Facility provides high quality and high throughput cryoEM data collection for HHMI users. The facility is not intended to spend a significant amount of Krios microscope time or staff hours for screening grids. Users of the facility should review policies below and know how to best utilize their time during their scheduled sessions.

Screening cryo grid quality

We strongly recommend users to screen their cryo grids prior to shipping their grids to the Janelia CryoEM facility. Please screen 1 or 2 grids from the same sample batch to assess ice coverage, ice contamination and particle concentration. Upon shipping grids, please fill in the Grid submission form (https://www.cognitoforms.com/HHMI5/HHMICryoEMGridSubmission) and upload screened images if available. If we determine the user’s grids do not have particles or enough targets for data collection, the Janelia CryoEM Facility reserves the right to cancel the session and charge the user for the remainder of their session. 

Preparing and shipping cryo grids

We strongly encourage users to clip their grids into autogrids before shipping. This will save sample handling and loading time on the first day of the scheduled data collection session; thus, the user will have more time for data acquisition.

Cryo grids shipped to us should be stored in the least number of cryo boxes (for example, 8 cryo grids should be stored in 2 cryo boxes instead of 3 or more boxes). The cryo boxes should be clearly labeled. Users should choose cryoboxes that can be loosened with a commonly available tool such as a small Philip or slot screwdriver. Please do not overtighten the lids on the cryo box or on the Falcon tube. 

After loading the grids, aligning and calibrating the microscope and camera, our staff will assist users to check a limited number of grids. Due to limited standard work hours, we will not guarantee to screen all the loaded grids on a given day.

Users should ship their cryo grids to the Janelia CryoEM Facility 1-5 business days before their scheduled session. Grids may be personally delivered to the Janelia CryoEM Facility if the grids arrive on the morning of the first day of their scheduled session.

Our shipping address and detailed instructions can be found on the PDF below. (https://www.janelia.org/sites/default/files/Support%20Teams/Cryo-Electron/instruction-on-shipping-cryogrids-Jan2020.pdf)

Please submit the grid submission form when you ship grids.(https://www.cognitoforms.com/HHMI5/HHMICryoEMGridSubmission) 

If users wish to save grids for future sessions, we can store the grids for up to 2 months upon request.

Data collection

To best use resources at the Janelia CryoEM facility, users should fill in a priority list of grids on the grid submission form and be available to communicate with our staff while setting up data acquisition. Our staff will check a limited number of grids and assess grid quality for users, but users are ultimately responsible for deciding which grid to set up automated data collection before 3:00pm Eastern Time. Users may continue to screen their grids after 3:00pm Eastern Time but will lose their opportunity to set up overnight data collection that day.

To ensure the optimal performance of the microscope and to minimize microscope downtime, Janelia CryoEM Facility staff operate and align the microscope, calibrate the camera and optimize the imaging condition and parameters for each data collection session. We typically set up automated data collection in SerialEM for a user with the user picking regions of interest (ROIs) for the collection. If a user is well experienced with SerialEM data collection and desires to set up the data collection themselves, they first need to work one session under the supervision of a staff member to obtain permission for setting up data collection in future sessions. Users are not permitted to align the microscope, change major settings on the scope/camera or load a new cassette of grids without consulting with a staff member. If a user setting up data collection on their own needs to change grids, they should do so during standard work hours when a Janelia staff member is around in case unexpected issues occur. If a hardware/software issue occurs during data collection, a user should not try to solve the issue themselves without consulting with a staff member. 

Data transfer and storage

Users may use Globus or mail us an external hard drive to transfer collected data. Due to limited resources, we can only store user’s data for up to 2 months. 

Publication policy

For routine data collection services on user-provided ready-to-image cryo grids (Janelia CryoEM facility does not help on grids preparation or image processing but only use standard data collection schemes to collect data  for users), the following acknowledgment should be included in the publication: 1, When describing data collection in the Materials and Methods or equivalent section, please mention that data was collected on a 300 kV FEI Titan Krios microscope located at the HHMI Janelia Research Campus. 2, In the Acknowledgement section of the paper, please acknowledge help from the Janelia CryoEM facility staff. Here is one possible example: We thank staff name, staff name and staff name at the HHMI Janelia CryoEM Facility for help in microscope operation and data collection.

For non routine data collection sessions that Janelia CryoEM staff enabled customization or made significant intellectual input or overcame extra difficulties to make the work possible, they should be considered to be listed as co-authors.

Any projects where Janelia CryoEM staff prepare samples/grids, or carry out data processing are considered as collaborations.  Cryo-ET projects involving cryo FIB milling at Janelia CryoEM Facility should be considered as collaboration as significant amount of staff expertise and intellectual inputs are provided by the facility. For collaboration work that results in publication, CryoEM staff members should be listed as co-authors.