<div dir="ltr">To order O-rings, you may contact Tom. He repairs Gatan stuff. I once bot some O-rings from him. He is also our goto if anything wrong with out of warranty Gatan thing. Do check with Gatan first.<div><br></div><div><p class="MsoNormal" style="margin:0in 0in 0.0001pt 48pt;font-size:12pt;font-family:Calibri,sans-serif;color:rgb(0,0,0)"><span style="font-size:11pt">Tom Schmelzer</span></p><p class="MsoNormal" style="margin:0in 0in 0.0001pt;font-size:12pt;font-family:Calibri,sans-serif;color:rgb(0,0,0)"><b><span style="font-size:11pt;color:rgb(192,0,0)">TGS Technologies LLC</span></b><span style="font-size:11pt"></span></p><p class="MsoNormal" style="margin:0in 0in 0.0001pt;font-size:12pt;font-family:Calibri,sans-serif;color:rgb(0,0,0)"><b><span style="font-size:11pt;color:rgb(192,0,0)">USA</span></b><span style="font-size:11pt"></span></p><p class="MsoNormal" style="margin:0in 0in 0.0001pt;font-size:12pt;font-family:Calibri,sans-serif;color:rgb(0,0,0)"><span style="font-size:11pt">724-453-3865</span></p><p class="MsoNormal" style="margin:0in 0in 0.0001pt;font-size:12pt;font-family:Calibri,sans-serif;color:rgb(0,0,0)"><span style="font-size:11pt">email: <span style="color:rgb(149,79,114)"><a href="mailto:tom@tgstechnologies.net" title="mailto:tom@tgstechnologies.net" style="color:purple">tom@tgstechnologies.net</a></span></span></p><p class="MsoNormal" style="margin:0in 0in 0.0001pt;font-size:12pt;font-family:Calibri,sans-serif;color:rgb(0,0,0)"><span style="font-size:11pt"><br></span></p><p class="MsoNormal" style="margin:0in 0in 0.0001pt;font-size:12pt;font-family:Calibri,sans-serif;color:rgb(0,0,0)"><br></p></div></div><br><div class="gmail_quote"><div dir="ltr" class="gmail_attr">On Mon, Feb 24, 2020 at 4:01 PM Denis Chrétien <<a href="mailto:denis.chretien@univ-rennes1.fr">denis.chretien@univ-rennes1.fr</a>> wrote:<br></div><blockquote class="gmail_quote" style="margin:0px 0px 0px 0.8ex;border-left:1px solid rgb(204,204,204);padding-left:1ex"><div style="overflow-wrap: break-word;">If we order O-rings from Thermo, it cost one arm… Does anyone have references for « standard » O-rings that we can buy elsewhere for a much cheaper price?<div><br></div><div>To come back to the original questions, we still use 626 and other side-entry holders. Sometime we face vacuum problems. In our experience these are mainly due to (as already said):</div><div><br></div><div>- Dusts on the O-ring (or inside the airlock system). Inspect the O-ring and also try to blow dry-air inside the airlock. Maybe the airlock tube is damage, then ask Thermo for repair … :-(</div><div><br></div><div>- Dry O-rings: regularly put some (not a lot !) vacuum grease on it. Be careful not to scratch the metal when removing it or damaging the O-ring. Change it if dry or obviously damaged.</div><div><br></div><div>- One frequent issue with cryo-holders: do not fill the cryo-holder station with to much LN2, you may freeze the O-ring and it will inevitably lead to a vacuum leak (very reproducible!). To limit this issue, we slightly lift the cryo-holder station with a rectangular grid box to avoid LN2 going into contact with the O-ring. We (not every one of us) cool the cryo-stage inside the microscope. Then the work-station is cooled just before inserting the specimen grid on the holder. This also limits the risk of cooling the O-ring.</div><div><br></div><div>- We « pre-pump » the airlock, and pump for 1 min before inserting (on a Tecnai G2 Sphera). 3 min of pumping seams a lot!</div><div><br></div><div>- Change of the zeolithe, grease on the valve O-ring of the cryo-holder, periodic zeolithe cycle (we do it every session): it improves stability on our side.</div><div><br></div><div>Best</div><div><br></div><div>Denis</div><div><div><br><blockquote type="cite"><div>Le 24 févr. 2020 à 21:26, David Stokes <<a href="mailto:stokes@nyu.edu" target="_blank">stokes@nyu.edu</a>> a écrit :</div><br><div><span style="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;text-decoration:none;float:none;display:inline">I used to do the following prior to every session:</span><br style="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;text-decoration:none"><span style="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;text-decoration:none;float:none;display:inline">Remove o-ring from holder.</span><br style="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;text-decoration:none"><span style="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;text-decoration:none;float:none;display:inline">Clean o-ring off with kimwipes and fingers (I didn't bother to use any solvent).</span><br style="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;text-decoration:none"><span style="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;text-decoration:none;float:none;display:inline">Clean o-ring groove with kimwipes.</span><br style="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;text-decoration:none"><span style="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;text-decoration:none;float:none;display:inline">Blow away fibers with compressed air.</span><br style="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;text-decoration:none"><span style="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;text-decoration:none;float:none;display:inline">Lightly grease o-ring.</span><br style="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;text-decoration:none"><span style="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;text-decoration:none;float:none;display:inline">Inspect o-ring and o-ring groove under dissecting scope and carefully remove any fibers prior to re-installation onto the holder.</span><br style="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;text-decoration:none"><span style="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;text-decoration:none;float:none;display:inline">Don't scratch the o-ring groove. In fact, don't touch it with forceps. Use compressed air.</span><br style="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;text-decoration:none"><br style="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;text-decoration:none"><span style="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;text-decoration:none;float:none;display:inline">During the inspection process, you will see whether there are any nicks or flat spots in the o-ring. If so, then consider a replacement. If there is a scratch in the o-ring groove, then you might need a new holder (or maybe not).</span><br style="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;text-decoration:none"><span style="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;text-decoration:none;float:none;display:inline">Also make sure you really have the right sized o-ring. At some point in ancient history, they changed the size slightly, such that CM200 o-ring was a bit fatter than the one for the CM12 stage.</span><br style="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;text-decoration:none"><br style="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;text-decoration:none"><span style="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;text-decoration:none;float:none;display:inline">David</span><br style="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;text-decoration:none"><br style="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;text-decoration:none"><br style="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;text-decoration:none"><span style="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;text-decoration:none;float:none;display:inline">On 2/24/2020 3:10 PM, Sharon G Wolf wrote:</span><br style="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;text-decoration:none"><blockquote type="cite" style="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;text-decoration:none">Terje<br>We also have suffered for years from vacuum breaks. We tried EVERYTHING. The most effective thing was to (believe it or not) change the o-ring every single session.<br>Recently we understood this was due to the fibers and other "dirt" that we observed on the o-rings every time we looked with the binocular after inserting and removing the holder from the microscopes, transfer stations or pumping station. We embarked on a “cleaning program” where we gently swab all the above mentioned devises. It’s now slowly getting better. We are still changing the o rings but we hope it will soon no longer be necessary, once all the "stuff" that accumulated over the years is cleaned out from those places.<br>Sharon<br><br><br>Sent from my iPhone<br><br><blockquote type="cite">On 24 Feb 2020, at 19:28, Dokland, Terje <<a href="mailto:dokland@uab.edu" target="_blank">dokland@uab.edu</a>> wrote:<br><br>We have also had persistent vacuum break problems with our 626 holder in our Tecnai F20 over the past two years. About 50% of the time inserting the holder breaks the vacuum. I though it was unique to this holder, but with the Elsa holder that we recently acquired it is even worse: vacuum break about 90% if the time. We’ve changed O rings, had the airlock disassembled and cleaned, but the problem persists.<br><br>That being said, it never affected ice quality, and the temperature usually doesn’t go above -165 C. When the vacuum recovers it’s usually at 25-30 log or so and back to <10 within a half hour. It’s really annoying though.<br><br>In my experience as long as the temperature doesn’t rise above -150C you should be ok.<br><br>It helps to make sure the tip is very cold by submerging it completely in lN2 before inserting.<br><br>Terje<br><br>Sent from my iPhone<br><br><blockquote type="cite">On Feb 24, 2020, at 7:13 AM, TALAVERA PEREZ Ariel <<a href="mailto:Ariel.Talavera@ulb.be" target="_blank">Ariel.Talavera@ulb.be</a>> wrote:<br><br>Dear all,<br><br>I am using a Gatan 626 holder on a side entry TALOS microscope. I am<br>getting problems with very frequent crystalline ice formation on the<br>grids. Some times I get a mix of vitreous and crystalline water, but<br>must of the time I only get crystalline water. I have been extra careful<br>during sample preparation, and transferring the grind on to the holder,<br>and I have done 48 hours holder bake out before insertion, but I still<br>get ice on the grid.<br><br>I monitored the temperature of the holder during the insertion process,<br>including 3 minutes for the Airlock pump. During those 3 minutes the<br>temperature rises up to -152.2. Can this be the reason of the unwanted<br>ice formation? How high can be the temperature of the holder to ensure a<br>nice vitreous water?<br><br>I am also getting vacuum problems while inserting the holder. This<br>process has resulted to be extremely, extremely sensitive. After the 3<br>minutes Airlock pumping (I cannot give longer time because of the<br>aforementioned temperature issue) as soon as I open the column port the<br>vacuum either rises to 99 Log or completely crashes the column vacuum.<br>This has happened with two different holders either cooled with liquid<br>nitrogen or at room temperature. So, my question: Is this insertion<br>process always that picky or it can be that there is something wrong<br>with the holders/Airlock pump?<br><br>Thanks a lot in advanced.<br><br>Best regards.<br><br>Ariel<br><br>--<span> </span><br>Ariel TALAVERA PEREZ, PhD<br>Center for Microscopy and Molecular Imaging (CMMI)<br>Laboratory of Microscopy<br>Université Libre de Bruxelles<br>Rue Adrienne Bolland, 8<br>B-6041 Gosselies, Belgium<br>Phone +32 (0)26509866<br><br>_______________________________________________<br>3dem mailing list<br><a href="mailto:3dem@ncmir.ucsd.edu" target="_blank">3dem@ncmir.ucsd.edu</a><br><a href="https://urldefense.proofpoint.com/v2/url?u=https-3A__mail.ncmir.ucsd.edu_mailman_listinfo_3dem&d=DwIGaQ&c=slrrB7dE8n7gBJbeO0g-IQ&r=hgbDG_ZdQaorZsucl01zHg&m=Yn2a7P10qylNPjdZ499rPbNzetpn8RyAV870SJbr0hg&s=wwZt0ruHRcbRcKBOUphvqWKC9QI8lDUPt_L8YiYwln4&e=" target="_blank">https://urldefense.proofpoint.com/v2/url?u=https-3A__mail.ncmir.ucsd.edu_mailman_listinfo_3dem&d=DwIGaQ&c=slrrB7dE8n7gBJbeO0g-IQ&r=hgbDG_ZdQaorZsucl01zHg&m=Yn2a7P10qylNPjdZ499rPbNzetpn8RyAV870SJbr0hg&s=wwZt0ruHRcbRcKBOUphvqWKC9QI8lDUPt_L8YiYwln4&e=</a><br></blockquote>_______________________________________________<br>3dem mailing list<br><a href="mailto:3dem@ncmir.ucsd.edu" target="_blank">3dem@ncmir.ucsd.edu</a><br><a href="https://urldefense.proofpoint.com/v2/url?u=https-3A__mail.ncmir.ucsd.edu_mailman_listinfo_3dem&d=DwIGaQ&c=slrrB7dE8n7gBJbeO0g-IQ&r=hgbDG_ZdQaorZsucl01zHg&m=Yn2a7P10qylNPjdZ499rPbNzetpn8RyAV870SJbr0hg&s=wwZt0ruHRcbRcKBOUphvqWKC9QI8lDUPt_L8YiYwln4&e=" target="_blank">https://urldefense.proofpoint.com/v2/url?u=https-3A__mail.ncmir.ucsd.edu_mailman_listinfo_3dem&d=DwIGaQ&c=slrrB7dE8n7gBJbeO0g-IQ&r=hgbDG_ZdQaorZsucl01zHg&m=Yn2a7P10qylNPjdZ499rPbNzetpn8RyAV870SJbr0hg&s=wwZt0ruHRcbRcKBOUphvqWKC9QI8lDUPt_L8YiYwln4&e=</a><br></blockquote>_______________________________________________<br>3dem mailing list<br><a href="mailto:3dem@ncmir.ucsd.edu" target="_blank">3dem@ncmir.ucsd.edu</a><br><a href="https://urldefense.proofpoint.com/v2/url?u=https-3A__mail.ncmir.ucsd.edu_mailman_listinfo_3dem&d=DwIGaQ&c=slrrB7dE8n7gBJbeO0g-IQ&r=hgbDG_ZdQaorZsucl01zHg&m=Yn2a7P10qylNPjdZ499rPbNzetpn8RyAV870SJbr0hg&s=wwZt0ruHRcbRcKBOUphvqWKC9QI8lDUPt_L8YiYwln4&e=" target="_blank">https://urldefense.proofpoint.com/v2/url?u=https-3A__mail.ncmir.ucsd.edu_mailman_listinfo_3dem&d=DwIGaQ&c=slrrB7dE8n7gBJbeO0g-IQ&r=hgbDG_ZdQaorZsucl01zHg&m=Yn2a7P10qylNPjdZ499rPbNzetpn8RyAV870SJbr0hg&s=wwZt0ruHRcbRcKBOUphvqWKC9QI8lDUPt_L8YiYwln4&e=</a><br></blockquote><br style="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;text-decoration:none"><span style="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;text-decoration:none;float:none;display:inline">--<span> </span></span><br style="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;text-decoration:none"><span style="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;text-decoration:none;float:none;display:inline">David L. 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style="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;text-decoration:none"><a href="mailto:3dem@ncmir.ucsd.edu" style="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" target="_blank">3dem@ncmir.ucsd.edu</a><br style="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;text-decoration:none"><a href="https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem" style="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" target="_blank">https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem</a></div></blockquote></div><br></div></div>_______________________________________________<br>
3dem mailing list<br>
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<a href="https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem" rel="noreferrer" target="_blank">https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem</a><br>
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