National Cancer Institute - Cancer.gov

Cryo-EM Facility: User Policies

Procedures for Accessing the CSB Cryo-EM Facility

  1. Titan Krios G4 user sitting at workstation and looking at monitors
    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 (susan.lea@nih.gov), for approval and cc: Dan Shi (dan.shi@nih.gov) for documentation.
  2.  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.
  3. 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.
  4. 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
  5. 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).
  6. 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.