CryoEM Training

 

cryoEM training schedule

After finishing the cryoEM training, it is required to have 10 more supervised sessions on the microscope with your own sample untile you can pass the final test. Facility team will work with you until you get a satisfactory dataset.

For a detailed description of the training plan and schedule please go to this updating google docs: Purdue_cryoEM_training

For CM200 operation procedure:  CM200_manual  , will be replaced by Talos F200C in Feburary,2018.

For CM200 test: CM200_check_out

The service policy and training policy is changed, The Purdue Cryo-Electron Microscopy Facility (PCEMF) provides cryoEM training for those who have already completed the basic TEM training (TEM operation for room temperature tasks and preparation of negative stain samples) at the Purdue Electron Microscopy Facility (PEMF).  The cryoEM facility will not accept a non-cryoEM application. For the regular EM checking request, it's better to apply for the service from EM facility, so that the staff in EM Facility will help you for your specific project. Please spend one more hour on this page carefully, you might learn enough how we work on cryoEM project. This's not a simple equipment,  but a complicated system. You might spend at least two months on learning and practising. The most important thing is the PRACTICE.  There was no way to operate them well just by watching. I welcome you take your own samples for cryoEM training, and I am willing to introduce some tips for your specific project. Feel free email to us if you have any questions.
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For basic TEM training, please contact the Purdue Electron Microscopy Facility (PEMF):

Christopher J Gilpin, Ph.D.
Phone: (765) 494-7750
Email: gilpin@purdue.edu

You should ask training on T20 and T12 for screening negative staining grids. 

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For Cryo-EM training, please arrange a meeting with the following contact, complete the Purdue University Laboratory Safety Fundamentals (LSF) online training: http://purdue.vividlms.com/, and bring the  Certificate of Completion with you to the first training session.

Kunpeng Li,  Ph.D.
Hock 111 
Phone: (765) 421-5516
Email: li2182@purdue.edu
 
Available time for training and practice: 1:00pm~5:00pm (Tuesday - Friday)

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Cost comparison between training and purchasing full service

Please take a moment to consider the cost issue. If you only have one specific project with cryoEM, it’s cheaper to purchase a full service session, which would cost significantly less time and money. If you anticipate your project will require more than a few sessions on the electron microscope it would be wise to receive full training and certification on the instruments. For the full training, you might need to book 12 hours training sessions totally in one month and 10 more hours practice sessions with your sample under supervising.

What CryoEM Training focus on:

PCEMF is asking that everyone send us example micrographs of their good ice and good particles prior to making reservations on the Titan. Based on this rule, Our training will focus on how to screen your grids and samples, besides on equipment operation and risk control issues. You will go through all the basic details about how to freeze a perfect grid for the final data collection on Titan.

CryoEM training tutorials (In order to carry out a research program)

1) The basic knowledge and procedures of CryoEM (Vanessa Cabra, Montserrat Samsó)

2) From Grid Preparation to Image Acquisition (David S. Booth, Agustin Avila-Sakar, Yifan Cheng)

3) Glow discharging grids with the Emitech K950x(Brenda in Wen Jiang group):

4) Sample qualification: the optimized protocol for asymmetric small proteins (Ren Gang)

5) How to make a GO grid(Thomas G. Martin in Sjors H. W. Scheres group)

Titan training (only for advanced users)

We don't have a dedicated training on Titan, The staffs will do training when you collect real data on Titan. For the senior cryoEM users, there are additional training options including instructions on:

  • Autogrid assembly

  • Load autogrids on cassette

  • Load cassette on autoloader

  • Direct alignment, objective lens stigmator and comma free alignment

  • K2 camera and DM software

  • Titan column/autoloader conditioning and K2 cryo-cycle

  • Leginon data collection and Appion data processing

For a new user, personally I suggest you come to watch the staff or senior students who work on Titan when they do alignment or run Leginon. According to my experiences, it will take some time to learn everything well, so it will be necessary to watch several times.


But, We don't have a dedicated training on Titan, Valorie and Thomas will do training when you collect real data on Titan.ButButPersonally I suggest you assign a  student to watch the staff or senior students who work on Titan when they do alignment or run Leginon. You can get the schedule plan from Valorie, just feel free to contact the staff or senior students who will be responsible for the specific Titan sessions.  To my experiences, it will take some time to learn everything well, so it will be
necessary to watch several times.