<html><head><meta http-equiv="Content-Type" content="text/html; charset=utf-8"></head><body style="word-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;" class="">Hi Bradford,<div class=""><br class=""></div><div class="">We use serial EM on a tecnai T12 LaB6 TEM equipped with an 4K eagle for SPA. We run the software through windows XP without problem. You only need to have enough memories to run the program. We used it for negative stain and cryo EM data acquisition. You can easily do low dose imaging and the software also provides tomography acquisition which is sometimes better than the FEI software. Installation of the software is easy and will take you only 3-4hours. On the website, the directives for installation are clear and show you step by step how to install the software.</div><div class=""><br class=""></div><div class="">The resolution that you will obtain with a 200 can be pretty good and you will be able to publish novel data with it but the use of an Artica equipped with a direct electron camera will surely improve the resolution. </div><div class=""> </div><div class="">Hope that will help you. </div><div class=""><br class=""></div><div class="">Regards,</div><div class=""><div class="">
<div dir="auto" style="caret-color: rgb(0, 0, 0); color: rgb(0, 0, 0); letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; text-decoration: none; word-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;" class=""><div dir="auto" style="caret-color: rgb(0, 0, 0); color: rgb(0, 0, 0); letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; text-decoration: none; word-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;" class=""><div dir="auto" style="text-align: start; text-indent: 0px; word-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;" class=""><div dir="auto" style="text-align: start; text-indent: 0px; word-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;" class=""><div dir="auto" style="text-align: start; text-indent: 0px; word-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;" class=""><div style="word-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;" class=""><div style="caret-color: rgb(0, 0, 0); color: rgb(0, 0, 0); letter-spacing: normal; text-transform: none; white-space: normal; word-spacing: 0px; text-decoration: none; -webkit-text-stroke-width: 0px; font-family: Helvetica; font-size: 12px; font-style: normal; font-variant-caps: normal; font-weight: normal; text-align: start; text-indent: 0px;">Aurélien FOUILLEN, PhD</div><div style="caret-color: rgb(0, 0, 0); color: rgb(0, 0, 0); letter-spacing: normal; text-transform: none; white-space: normal; word-spacing: 0px; text-decoration: none; -webkit-text-stroke-width: 0px; font-family: Helvetica; font-size: 12px; font-style: normal; font-variant-caps: normal; font-weight: normal; text-align: start; text-indent: 0px;">Post-doctorat<br class=""><br class=""></div><div style="caret-color: rgb(0, 0, 0); color: rgb(0, 0, 0); letter-spacing: normal; text-transform: none; white-space: normal; word-spacing: 0px; text-decoration: none; -webkit-text-stroke-width: 0px; font-family: Helvetica; font-size: 12px; font-style: normal; font-variant-caps: normal; font-weight: normal; text-align: start; text-indent: 0px;">Université de Montréal<br class="">Laboratoire du Dr Antonio NANCI<br class="">Faculté de Médecine Dentaire<br class="">Département de Stomatologie</div><div style="caret-color: rgb(0, 0, 0); color: rgb(0, 0, 0); letter-spacing: normal; text-transform: none; white-space: normal; word-spacing: 0px; text-decoration: none; -webkit-text-stroke-width: 0px; font-family: Helvetica; font-size: 12px; font-style: normal; font-variant-caps: normal; font-weight: normal; text-align: start; text-indent: 0px;">Département de Biochimie et de Médecine Moléculaire<br class="">2900 Boul Edouard Montpetit</div><div style="caret-color: rgb(0, 0, 0); color: rgb(0, 0, 0); letter-spacing: normal; text-transform: none; white-space: normal; word-spacing: 0px; text-decoration: none; -webkit-text-stroke-width: 0px; font-family: Helvetica; font-size: 12px; font-style: normal; font-variant-caps: normal; font-weight: normal; text-align: start; text-indent: 0px;">Pavillon Roger Gaudry- Local A209<br class=""><br class="">1-(514)-343-5231<br class=""><a href="mailto:aurelien.fouillen@umontreal.ca" class="">aurelien.fouillen@umontreal.ca</a></div><div style="text-align: start; text-indent: 0px;"><a href="https://nancilaboratory.com" class="">https://nancilaboratory.com</a></div><div style="caret-color: rgb(0, 0, 0); color: rgb(0, 0, 0); letter-spacing: normal; text-transform: none; white-space: normal; word-spacing: 0px; text-decoration: none; -webkit-text-stroke-width: 0px; font-family: Helvetica; font-size: 12px; font-style: normal; font-variant-caps: normal; font-weight: normal; text-align: start; text-indent: 0px;">============================================</div></div></div></div></div></div></div>
</div>
<div><br class=""><blockquote type="cite" class=""><div class="">Le 10 oct. 2018 à 17:35, Grassucci, Robert <<a href="mailto:rg2502@cumc.columbia.edu" class="">rg2502@cumc.columbia.edu</a>> a écrit :</div><br class="Apple-interchange-newline"><div class=""><div class="WordSection1" style="page: WordSection1; caret-color: rgb(0, 0, 0); font-family: Helvetica; font-size: 12px; font-style: normal; font-variant-caps: normal; font-weight: normal; letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; text-decoration: none;"><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);" class="">Hi Bradford,<o:p class=""></o:p></span></div><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);" class="">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<span class="Apple-converted-space"> </span><a href="http://bio3d.colorado.edu/SerialEM/" style="color: rgb(149, 79, 114); text-decoration: underline;" class="">http://bio3d.colorado.edu/SerialEM/</a><o:p class=""></o:p></span></div><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);" class="">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:</span><span class="Apple-converted-space"> </span><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);" class=""><a href="http://emg.nysbc.org/redmine/projects/leginon/wiki/Leginon_Homepage" style="color: rgb(149, 79, 114); text-decoration: underline;" class="">http://emg.nysbc.org/redmine/projects/leginon/wiki/Leginon_Homepage</a><o:p class=""></o:p></span></div><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);" class=""><o:p class=""> </o:p></span></div><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);" class="">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.<o:p class=""></o:p></span></div><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);" class="">Regards,<o:p class=""></o:p></span></div><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);" class="">Bob<o:p class=""></o:p></span></div><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);" class=""><o:p class=""> </o:p></span></div><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);" class=""><o:p class=""> </o:p></span></div><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);" class=""><o:p class=""> </o:p></span></div><div class=""><div style="border-style: solid none none; border-top-width: 1pt; border-top-color: rgb(225, 225, 225); padding: 3pt 0in 0in;" class=""><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><b class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif;" class="">From:</span></b><span style="font-size: 11pt; font-family: Calibri, sans-serif;" class=""><span class="Apple-converted-space"> </span>3dem [<a href="mailto:3dem-bounces@ncmir.ucsd.edu" style="color: rgb(149, 79, 114); text-decoration: underline;" class="">mailto:3dem-bounces@ncmir.ucsd.edu</a>]<span class="Apple-converted-space"> </span><b class="">On Behalf Of<span class="Apple-converted-space"> </span></b>Ross, Bradford<br class=""><b class="">Sent:</b><span class="Apple-converted-space"> </span>Wednesday, October 10, 2018 4:54 PM<br class=""><b class="">To:</b><span class="Apple-converted-space"> </span><a href="mailto:3dem@ncmir.ucsd.edu" style="color: rgb(149, 79, 114); text-decoration: underline;" class="">3dem@ncmir.ucsd.edu</a><br class=""><b class="">Subject:</b><span class="Apple-converted-space"> </span>[3dem] Setting up a 200kV Tecnai with 4K Eagle for SPA: Worth bothering?<o:p class=""></o:p></span></div></div></div><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><o:p class=""> </o:p></div><div class=""><div class=""><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 10pt; font-family: Tahoma, sans-serif;" class="">Hello 3DEM Community,<o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 10pt; font-family: Tahoma, sans-serif;" class=""><o:p class=""> </o:p></span></div></div><div class=""><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 10pt; font-family: Tahoma, sans-serif;" class="">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.<o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 10pt; font-family: Tahoma, sans-serif;" class=""><o:p class=""> </o:p></span></div></div><div class=""><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 10pt; font-family: Tahoma, sans-serif;" class="">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.<o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 10pt; font-family: Tahoma, sans-serif;" class=""><o:p class=""> </o:p></span></div></div><div class=""><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 10pt; font-family: Tahoma, sans-serif;" class="">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?<o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 10pt; font-family: Tahoma, sans-serif;" class=""><o:p class=""> </o:p></span></div></div><div style="margin: 0in 0in 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 10pt; font-family: Tahoma, sans-serif;" class="">Cheers,<o:p class=""></o:p></span></div><div class=""><div class=""><div class=""><p class="MsoNormal" style="margin: 0in 0in 12pt; font-size: 12pt; font-family: "Times New Roman", serif;"><span style="font-size: 10pt; font-family: Tahoma, sans-serif;" class="">Bradford Ross<br class=""><br class="">Electron Microscopy Technician<br class="">BioImaging Facility<br class="">University of British Columbia<br class="">Cunningham Building Rm. 64<br class="">2146 East Mall <span class="Apple-converted-space"> </span><br class="">Vancouver, B.C. <span class="Apple-converted-space"> </span><br class="">V6T 1Z4<br class=""><br class="">phone 604-822-6996<o:p class=""></o:p></span></p></div></div></div></div></div><span style="caret-color: rgb(0, 0, 0); font-family: Helvetica; font-size: 12px; font-style: normal; font-variant-caps: normal; font-weight: normal; letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; text-decoration: none; float: none; display: inline !important;" class="">_______________________________________________</span><br style="caret-color: rgb(0, 0, 0); font-family: Helvetica; font-size: 12px; font-style: normal; font-variant-caps: normal; font-weight: normal; letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; text-decoration: none;" class=""><span style="caret-color: rgb(0, 0, 0); font-family: Helvetica; font-size: 12px; font-style: normal; font-variant-caps: normal; font-weight: normal; letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; text-decoration: none; float: none; display: inline !important;" class="">3dem mailing list</span><br style="caret-color: rgb(0, 0, 0); font-family: Helvetica; font-size: 12px; font-style: normal; font-variant-caps: normal; font-weight: normal; letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; text-decoration: none;" class=""><a href="mailto:3dem@ncmir.ucsd.edu" style="color: rgb(149, 79, 114); text-decoration: underline; font-family: Helvetica; font-size: 12px; font-style: normal; font-variant-caps: normal; font-weight: normal; letter-spacing: normal; orphans: auto; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: auto; word-spacing: 0px; -webkit-text-size-adjust: auto; -webkit-text-stroke-width: 0px;" class="">3dem@ncmir.ucsd.edu</a><br style="caret-color: rgb(0, 0, 0); font-family: Helvetica; font-size: 12px; font-style: normal; font-variant-caps: normal; font-weight: normal; letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; text-decoration: none;" class=""><a href="https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem" style="color: rgb(149, 79, 114); text-decoration: underline; font-family: Helvetica; font-size: 12px; font-style: normal; font-variant-caps: normal; font-weight: normal; letter-spacing: normal; orphans: auto; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: auto; word-spacing: 0px; -webkit-text-size-adjust: auto; -webkit-text-stroke-width: 0px;" class="">https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem</a></div></blockquote></div><br class=""></div></body></html>