Scientific Facilities

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CryoElectron Microscopy lab

The cryoEM lab at NTU for imaging biological samples was set up to support research at LKCM and the School of Biological Sciences. The applications include single particle analysis of purified proteins and viruses, electron tomography of cell sections, bacteria and tissue samples, and using novel labeling techniques to image proteins inside the cell, and in isolation.

 

Imaging tools


  • Titan Krios 300kV cryo TEM equipped with energy filter, STEM, K2 and Falcon3EC detectors and Volta phase plate
  • Arctica 200kV cryo TEM equipped with Falcon3EC detector and Volta phase plate
  • T12 Icorr 120kV TEM for teaching, screening and correlative light and EM
  • Corrsight cryo confocal and spinning disk


Sample prep


  • Dual E-beam, carbon coaters and low-angle shadowing
  • Vitrobot for making cryo grids purified samples
  • Cryosectioning of cells
  • Apex2 and miniSOG labeling


Computing and image processing


  • Single particle analysis
  • Electron tomography and subtomogram averaging


 

Guidelines for access to NISB Cryo-Electron Microscopy facility


Initial User meeting & Training

  1. An initial user meeting involving the new user and a supervisor is a prerequisite to using the facility. The project assessment meeting with the Cryo-EM facility staff will help to identify suitable technique, instrument and training. The EM manager will go through the safety regulations, training requirements and instrument charges at this meeting.
  2. All users are required to submit an application form (T12/Arctica or Titan Krios). The supervisor/PI needs to sign the form and accept the terms and conditions of using Cryo-EM Facility. The Supervisor is responsible for authorizing the work and for payment of any costs incurred such as for training, consumables, equipment use or the cost of repairing equipment damaged as a result of incorrect operation by the User.
  3. For access to cryo-EM lab (irregardless of which microscope), all users are required to submit Office of Health & Safety (with risk assessment, sample declaration and safety acknowledgement form if needed) to EMicroscope@ntu.edu.sg before they are allowed to use the facility. The risk assessment is submitted online through the WRAS System (Click here).
  4. Only trained users are allowed to book and use instruments independently. New users are  required to schedule training with the Cryo-EM facility staff. Please complete and submit the NTU Cryo-EM lab training request.
  5. New users will be under the supervision of the Cryo-EM facility staff until till he/she becomes a proficient user.



Booking information and Rules (Except Titan Krios, see below)

  1. Instruments can be booked online (FOM) after registration and training.
  2. Only one session can be book in advance and the next session can only be booked upon completion of the previous session.
  3. Activities such as repairs and maintenance, undergraduate classes and training of new users are usually undertaken during normal business hours and occasionally this will significantly limit the time available for regular bookings.
  4. The user must attend every session you are booked for, unless there are exceptional circumstances beyond your control (such as illness). Charges will still apply if the user fail to turn up for bookings.
  5. A microscope can only be used by the person who booked it. Bookings are NOT transferable.
  6. If instrument time is available, the user may extend the microscope session outside the booked time (subjected to approval from Cryo-EM manager).
  7. Special experiments that may require long acquisition times are also granted based on a case by case basis. Please send in request through the Cryo-EM manager.



Titan Krios Usage


  1. Users applying for time on Titan Krios will need to submit a scientific proposal (Titan Krios).
  2. All applications will be evaluated by an evaluating committee and awarded time based on scientific merit. Users need to show that suitable cryo-EM samples have been produced for data collection.
  3. Once users has been awarded time, users may email the Cryo-EM facility staff to schedule time on Titan Krios. Only Cryo-EM staff can operate the Titan Krios.