FAQs
Home Cryo EM Course Time Reservation Info Protocols and Videos Training Useful Links FAQs
Q: How do I reserve time on a microscope?
Please refer to https://wiki.bnl.gov/LBMS/index.php/Time_Reservation_Info
Q: Do I need prior experience in cryo-EM sample prep or imaging?
Prior experience in cryo-EM sample imaging is preferable, but not necessary. We will provide you with the training and tools necessary for you to succeed in gathering data during your reservation.
Q: What training will I have to take?
Users must complete the following training modules before arriving on site:
https://training.bnl.gov/portal/GE-CYBERSEC
https://training.bnl.gov/portal/GE-COMPUSE-AGREE
https://training.bnl.gov/portal/TQ-GSO
https://training.bnl.gov/Portal/TQ-covid
Along with these safety trainings, users are asked to take the following instructor guided trainings on site.
Negative staining sample preparation
•Read Negative Staining Protocol
•Half day for up to three people
Cryo-sample preparation with Vitrobot
•Read Vitrobot Protocol
•One day for up to three people, and one more day to practice (preferred to have their own samples).
Talos operation with room temperature holder/sample
•One day for each person including sample loading/unloading, EM alignment, EPU for lacey/continuous carbon grid
•One day to review/practice withing two weeks
Talos operation with Cryo-sample/holder after training with room temperature holder/sample
• Read cryo-holder protocol
•A two-day session for each person including sample loading/unloading, EM alignment, EPU for holey carbon grids
•One day to review/practice within two weeks.
Q: How many days of in person training do I have to take?
The number of days vary depending on which in person training you will take, but please expect and plan to utilize about 5 days for training.
Q: How many days can I schedule a microscope for?
•Each GU proposal may have up to two reservations.
•Each BAG proposal may have up to four reservations
•Reservations may be made at most 3 months in advance
•Each reservation can be at most 6 shifts for high-end EM and 2 shifts for screening EMs, and at least 2 shifts for high-end EM.
•For each cycle, up to the allocated time may be reserved.
Q: Do I need to bring my own samples for training?
LBMS has samples on site to practice. You are welcome to bring your own samples for training and practice, but it is not necessary.
Q: Do I have to have my samples prepared before I arrive on site?
At LBMS we have the equipment necessary to prepare your cryo samples on site. You must take the required trainings before preparing your samples.
Q: How do I gain access to the BNL site?
You can find information on how to access the site at the LBMS homepage : LBMS Homepage
Q: Which grids are preferred? Which do you have available?
information in EM time reservation We have continuous grids along with 1.2/1.3 holey carbon grids.
Q: How do I ship my sample?
•A Safety Approval Form (SAF) must be submitted at least 14 days in advance ofyour scheduled EM time for remote experiments and 21 days in advance for proposals with onsite users. To ship samples to LBMS, you must have an approved SAF. Once approved, you can print a shipping label from the "General" tab on your SAF in PASS.
•To ensure a timely handling of the shipment, please use the shipping address shown in the “Dry Shipper Information”form ([1]), put a copy of the completed form and a FedEx return label inside the dry shipper, and email “FedEx Incoming” tracking numbers to your LBMS contact.
•Samples must arrive at LBMS at least one week prior to the allocated microscope time. If this is not possible, or for questions, please contact the LBMS local contact for your microscope time. Please always send in duplicate grids as accidents may happen.
•A completed sample form ([2]) must be submitted to lbms-em@bnl.govbefore sending samples to LBMS.
•A completed imaging parameters form ([3]) must be submitted to lbms-em@bnl.gov at least one week prior to the allocated microscope time.Note: Please see details at [4].
•Please note that failure to send the samples on time, or provide the dry-shipper form, sample form and imaging parameter form on time, can result in cancellation of your EM time.
Q: How do I prepare for data processing? What data processing software do you suggest?
Commonly used image processing softwares are available online:
Q: How can I transfer the data to my home institution?
we use Globus for secure file transfers from BNL to your home institution. Please find the tutorial for Globus account setup and file transfer at the following link: Globus Access Tutorial