<div dir="ltr">Hi Peter,<div><br></div><div>Whether it is because the clip ring didn't make good contact with grid or something else, the cryo-grid got warmed up inside the column and freeze dried due to the high vacuum.</div><div>We do see this kind of ice on both F20 and Polara from time to time, and believe it happened due the loose contact between grid and holder/clamp ring.</div><div><br></div><div>Best,</div><div><br></div><div>Zongli</div></div><br><div class="gmail_quote"><div dir="ltr">On Wed, Oct 17, 2018 at 10:32 AM <<a href="mailto:duncan.sousa@rcn.com">duncan.sousa@rcn.com</a>> wrote:<br></div><blockquote class="gmail_quote" style="margin:0 0 0 .8ex;border-left:1px #ccc solid;padding-left:1ex"><div><div style="font-family:times new roman,new york,times,serif;font-size:12pt;color:#000000"><div>Hi,</div><div><br></div><div>I agree. It looks like freeze dried protein solution. I'd definitely check the holder first, but if you can't find the problem there then check every stage that involves a grid transfer. For example, I've seen this happen when the LN2 level gets too low in a dewar and the grid sits in cold N2 gas for a while. <br></div><div><br></div><div>Duncan<br></div><br><hr id="m_2049180482311409351zwchr"><div style="color:#000;font-weight:normal;font-style:normal;text-decoration:none;font-family:Helvetica,Arial,sans-serif;font-size:12pt"><b>From: </b>"Mariena Silvestry Ramos" <<a href="mailto:ms3289@cornell.edu" target="_blank">ms3289@cornell.edu</a>><br><b>To: </b>"Eric Hanssen" <<a href="mailto:ehanssen@unimelb.edu.au" target="_blank">ehanssen@unimelb.edu.au</a>><br><b>Cc: </b><a href="mailto:3dem@ncmir.ucsd.edu" target="_blank">3dem@ncmir.ucsd.edu</a><br><b>Sent: </b>Tuesday, October 16, 2018 8:39:43 PM<br><b>Subject: </b>Re: [3dem] weird ice?<br><br>
Hi Peter,
<div><br>
</div>
<div>I second Eric. Looks like a bad contact (or lack of) between the clipring and the grid. This will cause the ice to devitrify and "crust" and it you park the beam, the stuff will fold back and do all sorts of weird moving/behaviour. I'd check if your clipping
mechanism is good. It could be that the wire that keeps the grid put in is worn, or, depending on the model, the ring is crossthreaded. Changing the clip wire is generally easy to do, with the right tools. I've done it a couple of times and it fixes the issue.
Definitely contact Gatan with any questions. Mark Dods and/or Braden Bowers are at the factory in Warrendale and can give you advice on what to test and the part(s) to purchase. </div>
<div><br>
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<div>Best of luck!</div>
<div><br>
</div>
<div>Mariena<br>
<br>
<div id="m_2049180482311409351AppleMailSignature" dir="ltr">Sent from my Game Boy</div>
<div dir="ltr"><br>
On Oct 16, 2018, at 8:15 PM, Eric Hanssen <<a href="mailto:ehanssen@unimelb.edu.au" target="_blank">ehanssen@unimelb.edu.au</a>> wrote:<br>
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<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1f497d">HI Peter,
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<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1f497d">They both look like the clip ring in your 626 (I am guessing that is the holder you are using) is not clipped properly, hence your vitreous ice has devitrified
and in the second picture it is even worse it has sublimed in the microscope and you have only the protein/salt scaffold left.
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<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1f497d">Regard</span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1f497d">Eric</span></p>
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<p class="MsoNormal"><b><span style="font-size:10.0pt;font-family:"Comic Sans MS";color:navy">-----------------</span></b></p>
<p class="MsoNormal"><b><span style="font-size:10.0pt;font-family:"Comic Sans MS";color:navy">Assoc. Prof Eric Hanssen</span></b></p>
<p class="MsoNormal"><b><span style="font-size:10.0pt;font-family:"Comic Sans MS";color:navy"> </span></b></p>
<p class="MsoNormal"><b><span style="font-size:10.0pt;font-family:"Comic Sans MS";color:#1f497d">Head - Advanced Microscopy Facility
</span></b><b><span style="font-size:10.0pt;font-family:"Comic Sans MS";color:#1f497d"></span></b></p>
<p class="MsoNormal"><b><span style="font-size:10.0pt;font-family:"Comic Sans MS";color:#1f497d">Honorary Principal Fellow – Department of Biochemistry and Molecular Biology</span></b></p>
<p class="MsoNormal"><b><span style="font-size:10.0pt;font-family:"Comic Sans MS";color:#1f497d"> </span></b></p>
<p class="MsoNormal"><b><span style="font-size:10.0pt;font-family:"Comic Sans MS";color:#1f497d">President Australian Microscopy and Microanalysis Society</span></b></p>
<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Comic Sans MS";color:#1f497d"><br>
Bio21 Molecular Science and Biotechnology Institute<br>
</span><span style="font-size:10.0pt;font-family:"Comic Sans MS";color:gray">30 Flemington Road - The University of Melbourne - Victoria 3010 - Australia<br>
email: </span><span style="font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1f497d"><a href="mailto:ehanssen@unimelb.edu.au" target="_blank"><span style="font-size:10.0pt;font-family:"Comic Sans MS";color:gray">ehanssen@unimelb.edu.au</span></a></span><span style="font-size:10.0pt;font-family:"Comic Sans MS";color:gray">
| Office: +61 3 83442449 | Microscope: +61 3 83442509 </span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Comic Sans MS";color:gray"> </span></p>
<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Comic Sans MS";color:gray"><image001.png> <image002.png></span></p>
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<p class="MsoNormal"><b><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif"" lang="EN-US">From:</span></b><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif"" lang="EN-US"> 3dem [<a href="mailto:3dem-bounces@ncmir.ucsd.edu" target="_blank">mailto:3dem-bounces@ncmir.ucsd.edu</a>]
<b>On Behalf Of </b>Peter Shen<br>
<b>Sent:</b> Wednesday, 17 October 2018 10:39 AM<br>
<b>To:</b> <a href="mailto:3dem@ncmir.ucsd.edu" target="_blank">3dem@ncmir.ucsd.edu</a><br>
<b>Subject:</b> [3dem] weird ice?</span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black">Hi everyone,
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black">We've recently started to see some funky ice on our TF20. Here are a couple of screenshots:</span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black"><a href="https://www.dropbox.com/sh/mlfado5rf9o447c/AAAt9VJM28KG6sCaNnCXIXQga?dl=0" target="_blank">https://www.dropbox.com/sh/mlfado5rf9o447c/AAAt9VJM28KG6sCaNnCXIXQga?dl=0</a></span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black">Has anyone seen this before? We see this regardless of sample and grid type (a few different protein preps and UltrAuFoil/Quantifoil tested). Any insight would
be greatly appreciated! We're pulling our hair out here... </span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black">Peter</span></p>
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