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

[Ross, Bradford]

Hello again 3DEM,

I just wanted to say thank you for the fast and helpful responses to my question. I have started digging into the documentation on the installation of SerialEM, and have already received a custom config framework from Guenter Resch as a starting point.

A few of you have chimed in with similar systems that are being used for screening, negative-stain SPA, and a bit of cryo use as well. Good to know that we can still generate some useful data, and I'm looking forward to expanding my skill set in this area.

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

________________________________
From: 3dem [3dem-bounces at ncmir.ucsd.edu] on behalf of Ross, Bradford [bradford.ross at botany.ubc.ca]
Sent: Wednesday, October 10, 2018 1:53 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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