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

[Tsybovsky, Yaroslav (NIH/NCI) [C]]

Hello Bradford,

We have a T20 with a LaB6 filament and an Eagle. You will have no problem running SerialEM on the microscope computer as long as you have the FEI scripting interface installed. Most systems do, and SerialEM website explains how to determine this. Setting up the software may take some time, from hours to days, depending on experience. The resolution will be limited by the thermionic gun (i.e., low spatial coherency and brightness) and the camera. This configuration will not be able to produce high-resolution (by modern standards) structures. Perhaps about 15 A is a realistic rough estimate. However, it will perform well as a screening microscope and for negative-stain single particle data collection.

Best regards,
Yaroslav

Yaroslav Tsybovsky, Ph.D. (Contractor)
Scientist II
Frederick National Laboratory for Cancer Research
Leidos Biomedical Research, Inc.
Post Office Box B
Frederick, Maryland 21702
Phone: 301-846-7550


From: Ross, Bradford <bradford.ross at botany.ubc.ca>
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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