<div dir="ltr"><div>Hi Bradford,</div><div><br></div><div>We have TF-20 that run on windows and have SerialEM on this PC for auto-data collection.</div><div>We use it mainly for negative stain SPA/RCT but it can be use easily with the 626 holder for cryo. <br></div><div><br></div><div>Best,<br></div></div><br><div class="gmail_quote"><div dir="ltr">On Wed, Oct 10, 2018 at 4:38 PM <<a href="mailto:3dem-request@ncmir.ucsd.edu">3dem-request@ncmir.ucsd.edu</a>> wrote:<br></div><blockquote class="gmail_quote" style="margin:0 0 0 .8ex;border-left:1px #ccc solid;padding-left:1ex">Send 3dem mailing list submissions to<br>
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Today's Topics:<br>
<br>
1. Setting up a 200kV Tecnai with 4K Eagle for SPA: Worth<br>
bothering? (Ross, Bradford)<br>
2. Re: Setting up a 200kV Tecnai with 4K Eagle for SPA: Worth<br>
bothering? (Grassucci, Robert)<br>
<br>
<br>
----------------------------------------------------------------------<br>
<br>
Message: 1<br>
Date: Wed, 10 Oct 2018 20:53:34 +0000<br>
From: "Ross, Bradford" <<a href="mailto:bradford.ross@botany.ubc.ca" target="_blank">bradford.ross@botany.ubc.ca</a>><br>
To: "<a href="mailto:3dem@ncmir.ucsd.edu" target="_blank">3dem@ncmir.ucsd.edu</a>" <<a href="mailto:3dem@ncmir.ucsd.edu" target="_blank">3dem@ncmir.ucsd.edu</a>><br>
Subject: [3dem] Setting up a 200kV Tecnai with 4K Eagle for SPA: Worth<br>
bothering?<br>
Message-ID:<br>
<<a href="mailto:B48A2601A4664A4AAF5A3D2F0E482801025E6088F4@s-itsv-mbx05p.ead.ubc.ca" target="_blank">B48A2601A4664A4AAF5A3D2F0E482801025E6088F4@s-itsv-mbx05p.ead.ubc.ca</a>><br>
Content-Type: text/plain; charset="iso-8859-1"<br>
<br>
Hello 3DEM Community,<br>
<br>
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.<br>
<br>
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.<br>
<br>
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?<br>
<br>
Cheers,<br>
Bradford Ross<br>
<br>
Electron Microscopy Technician<br>
BioImaging Facility<br>
University of British Columbia<br>
Cunningham Building Rm. 64<br>
2146 East Mall<br>
Vancouver, B.C.<br>
V6T 1Z4<br>
<br>
phone 604-822-6996<br>
<br>
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Message: 2<br>
Date: Wed, 10 Oct 2018 21:35:59 +0000<br>
From: "Grassucci, Robert" <<a href="mailto:rg2502@cumc.columbia.edu" target="_blank">rg2502@cumc.columbia.edu</a>><br>
To: "Ross, Bradford" <<a href="mailto:bradford.ross@botany.ubc.ca" target="_blank">bradford.ross@botany.ubc.ca</a>>,<br>
"<a href="mailto:3dem@ncmir.ucsd.edu" target="_blank">3dem@ncmir.ucsd.edu</a>" <<a href="mailto:3dem@ncmir.ucsd.edu" target="_blank">3dem@ncmir.ucsd.edu</a>><br>
Subject: Re: [3dem] Setting up a 200kV Tecnai with 4K Eagle for SPA:<br>
Worth bothering?<br>
Message-ID:<br>
<<a href="mailto:DM6PR02MB39454028563449FDBA41A8CA94E00@DM6PR02MB3945.namprd02.prod.outlook.com" target="_blank">DM6PR02MB39454028563449FDBA41A8CA94E00@DM6PR02MB3945.namprd02.prod.outlook.com</a>><br>
<br>
Content-Type: text/plain; charset="us-ascii"<br>
<br>
Hi Bradford,<br>
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 <a href="https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fbio3d.colorado.edu%2FSerialEM%2F&data=02%7C01%7Celad.binshtein%40vanderbilt.edu%7Cb93a5af55cf848d7e91408d62ef8c38a%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C0%7C636748043312411139&sdata=ggWd8%2F5hWDWWTitGU2LFZUKo3ZxF8TmBNmF53rXijDE%3D&reserved=0" rel="noreferrer" target="_blank">https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fbio3d.colorado.edu%2FSerialEM%2F&data=02%7C01%7Celad.binshtein%40vanderbilt.edu%7Cb93a5af55cf848d7e91408d62ef8c38a%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C0%7C636748043312411139&sdata=ggWd8%2F5hWDWWTitGU2LFZUKo3ZxF8TmBNmF53rXijDE%3D&reserved=0</a><br>
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: <a href="https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Femg.nysbc.org%2Fredmine%2Fprojects%2Fleginon%2Fwiki%2FLeginon_Homepage&data=02%7C01%7Celad.binshtein%40vanderbilt.edu%7Cb93a5af55cf848d7e91408d62ef8c38a%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C0%7C636748043312411139&sdata=snVbeaFaby03yXXy3HwtdGQ7pImeBc5VbECGWBlIguA%3D&reserved=0" rel="noreferrer" target="_blank">https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Femg.nysbc.org%2Fredmine%2Fprojects%2Fleginon%2Fwiki%2FLeginon_Homepage&data=02%7C01%7Celad.binshtein%40vanderbilt.edu%7Cb93a5af55cf848d7e91408d62ef8c38a%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C0%7C636748043312411139&sdata=snVbeaFaby03yXXy3HwtdGQ7pImeBc5VbECGWBlIguA%3D&reserved=0</a><br>
<br>
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.<br>
Regards,<br>
Bob<br>
<br>
<br>
<br>
From: 3dem [mailto:<a href="mailto:3dem-bounces@ncmir.ucsd.edu" target="_blank">3dem-bounces@ncmir.ucsd.edu</a>] On Behalf Of Ross, Bradford<br>
Sent: Wednesday, October 10, 2018 4:54 PM<br>
To: <a href="mailto:3dem@ncmir.ucsd.edu" target="_blank">3dem@ncmir.ucsd.edu</a><br>
Subject: [3dem] Setting up a 200kV Tecnai with 4K Eagle for SPA: Worth bothering?<br>
<br>
Hello 3DEM Community,<br>
<br>
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.<br>
<br>
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.<br>
<br>
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?<br>
<br>
Cheers,<br>
Bradford Ross<br>
<br>
Electron Microscopy Technician<br>
BioImaging Facility<br>
University of British Columbia<br>
Cunningham Building Rm. 64<br>
2146 East Mall<br>
Vancouver, B.C.<br>
V6T 1Z4<br>
<br>
phone 604-822-6996<br>
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End of 3dem Digest, Vol 134, Issue 19<br>
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</blockquote></div><br clear="all"><br>-- <br><div dir="ltr" class="gmail_signature" data-smartmail="gmail_signature"><div dir="ltr"><div><div dir="ltr"><div><div dir="ltr"><div><div dir="ltr"><div><div dir="ltr"><div><div dir="ltr"><div><div dir="ltr"><div><div dir="ltr"><div><div dir="ltr"><div><div dir="ltr"><div><div dir="ltr"><div style="background-color:rgb(255,255,255)" dir="ltr"><div dir="ltr" style="font-size:12.8px"><span style="font-size:12.8px">________________________________</span><font size="2"><br></font></div><div dir="ltr"><font size="2"><span style="background-color:rgb(255,255,255)">Elad Binshtein, Ph.D.</span></font></div><div dir="ltr"><font size="2">Cryo-EM specialist - Senior Staff Scientist<br style="background-color:rgb(255,255,255)"><span style="background-color:rgb(255,255,255)">Center for Structure Biology (CSB)</span><br>MCN Room 1207<br style="background-color:rgb(255,255,255)"><span style="background-color:rgb(255,255,255)">Vanderbilt University</span><br style="background-color:rgb(255,255,255)"><span style="background-color:rgb(255,255,255)">Nashville, TN</span><br>Office: +1-615-322-4671<br style="background-color:rgb(255,255,255)"><span style="background-color:rgb(255,255,255)">Mobile: <span><span><span><span>+1-615-481-4408</span><span dir="ltr"><span dir="ltr"><span><span></span></span></span></span></span></span></span></span><span style="background-color:rgb(255,255,0)"><span></span></span><br><a href="https://www.linkedin.com/in/elad-binshtein-6bb89428/" target="_blank">https://www.linkedin.com/in/elad-binshtein-6bb89428/</a></font></div><font style="background-color:rgb(255,255,0)" size="2"><span style="background-color:rgb(255,255,255)">E-Mail: </span><a style="background-color:rgb(255,255,255)" href="mailto:eladbi@gmail.com" target="_blank">elad.binshtein@vanderbilt.edu</a><br></font></div><div style="background-color:rgb(255,255,255)" dir="ltr"><span><div dir="ltr"><font size="2">twitter @EladBinshtein</font></div></span><font style="background-color:rgb(255,255,0)" size="2"></font></div><div><span style="font-size:12.8px">________________________________</span><br></div></div></div></div></div></div></div></div></div></div></div></div></div></div></div></div></div></div></div></div></div></div></div>