[3dem] Setting up a 200kV Tecnai with 4K Eagle for SPA: Worth bothering?

[Grassucci, Robert]

Hi Bradford,
The 2 data collection software that are most broadly used for SPA Leginon and SerialEM can be set up with a microscope running windows.  I have seen SerialEM http://bio3d.colorado.edu/SerialEM/
set up directly on a Tecnai running windows or on a separate PC controlling the camera and communicating with the microscope PC using a client program.  Leginon is generally run on a separate Linux computer and communicates with the microscope using a client program as well but requires a bit more to set up since it is linked to a database.  The particulars of that system are at: http://emg.nysbc.org/redmine/projects/leginon/wiki/Leginon_Homepage

Either system will work well for what you want to do but both require a bit of time to become comfortable with them  Gatan (Latitude) and Thermofisher (EPU) also sell SPA programs but I am not sure if they will work with an eagle camera.  The commercial systems may have a bit more direct support as you pay for it but both the SerialEM and Leginon folks are very quick to respond to any query.  Good luck.
Regards,
Bob



From: 3dem [mailto:3dem-bounces at ncmir.ucsd.edu] On Behalf Of Ross, Bradford
Sent: Wednesday, October 10, 2018 4:54 PM
To: 3dem at ncmir.ucsd.edu
Subject: [3dem] Setting up a 200kV Tecnai with 4K Eagle for SPA: Worth bothering?

Hello 3DEM Community,

We have a 200kV Tecnai G2 LaB6 TEM equipped with an Eagle 4K camera, Gatan 626 cryo holder, and a Vitrobot Mk IV. This instrument gets used fairly routinely for Cryo TEM of lipid nanoparticles, ambient temp TEM Tomography, and HRTEM of various types of nanoparticles down to approx. 1.4A crystal lattice resolution.

Lately I've been thinking about setting the system up to acquire single particle cryo EM data to expand our capabilities. However, I know that the newer systems such as Talos Arctica, Glacios, Krios etc. with direct electron detectors are far more capable than what we have. Further, our Tecnai is currently completely running in a Windows environment, so that complicates things a bit with the SPA software packages all running on Linux. Are there folks out there running similar systems for this purpose? Do you set the microscope PC up to dual boot with Linux for single particle acquisition, or use the support PC? I have no experience in single particle acquisition or analysis, but lots of experience doing low dose cryo TEM and tinkering with our lab equipment.

In short, would it be worth the effort of reconfiguring the system to acquire datasets that may be sub-par by today's standards? Or perhaps a useful hands-on experience in learning about how cryo SPA works, and expanding the capability of our lab?

Cheers,
Bradford Ross

Electron Microscopy Technician
BioImaging Facility
University of British Columbia
Cunningham Building Rm. 64
2146 East Mall
Vancouver, B.C.
V6T 1Z4

phone 604-822-6996
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