Procedures for Accessing the CSB Cryo-EM Facility
- susan.lea@nih.gov), for approval and cc: Dan Shi (dan.shi@nih.gov) for documentation. Submit a one-page proposal to state biological significance and current biochemistry, negative stain or cryo-EM result to the CSB lab chief, Dr. Susan Lea (
- After your project is approved, there are two ways to work on your project: 1) by your lab member(s); 2) by collaborating with a cryo-EM lab.
- If your lab has an expert with at least two years’ experience in the cryo-EM field, the expert user can access the Cryo-EM Facility to work on approved project(s). Evidence that expertise is in place for the full structure determination pipeline should be provided.
- Basic training on electron microscopy is open to people who have an approved project; they will be taught the basic concepts of electron optics and will be trained on the following aspects:
- Preparing negatively stained EM grids
- Operating the Talos L120C for screening negatively stained grids
- Using different plunge freezers for preparing and improving frozen-hydrated EM grids
- If you collaborate with any group(s) in the CSB, expert user(s) from those lab(s) will work through the whole procedure – from specimen preparation to data processing for/with your lab member(s).
- Cryo-EM expert training on high-throughput data collection and 3D reconstruction will be through a collaboration with cryo-EM labs in CSB or NIH.
How to Access Microscopes and Specimen Preparation Equipment
TFS Vitrobot and Leica EMGP: User can directly reserve a slot through CSB's booking calendar: http://bric.sbl.ncifcrf.gov/CEM/cryoem_instrument_calendars.html
Chameleon: Email Tapan Kanai (tapan.kanai@nih.gov) to request a training or freezing grids session.
Aquilos II: This is used only for Cryo-ET projects; email Dan Shi (dan.shi@nih.gov) to request training sessions.
Talos L120C: no EM experience user(s) should contact Dan Shi (dan.shi@nih.gov) for training sessions. Well trained users can find open slots from CSB's booking calendar and send an email to Dan Shi (dan.shi@nih.gov) for requesting available session(s) – maximum 2 hours per session.
Talos Arctica: Submit to Mi Li (lim@nih.gov) your grid sheet(s); he will schedule an Arctica session based on submitting time.
- Monday, Wednesday and Friday daytime for fast screening grids
- Tuesday, Thursday and overnights for a small set data collection which is for evaluating single particle quality
Krios G4: Send Dr. Susan Lea (susan.lea@nih.gov) a 3D map solved by data from the Arctica with an original proposal for approval, Justin Deme (justin.deme@nih.gov) will schedule microscope sessions.