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<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Hi Peter,<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">As people pointed out the crystalized ice should be caused by the grid’s temperature was raised before recording the images. Loosen clip ring usually associated longer settle
time and severe drifting. Extreme thing ice could be devitrified within a second in RT air as Tommi suggested, you could take an image from the thicker area of the grid to confirm it.<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Beside slightly increasing the ice thickness, shortening the grids travel distance/time between liquid ethane to LN2 during freezing may help too. I observed very similar incidents
of grids with relative thin ice on our Krios.<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Good luck,<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Dan Shi<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Frederick National Lab for Cancer research<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">CCR, NCI
<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"> <o:p></o:p></span></p>
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<p class="MsoNormal"><b><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">From:</span></b><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"> White, Tommi A. <whiteto@missouri.edu>
<br>
<b>Sent:</b> Wednesday, October 17, 2018 1:03 PM<br>
<b>To:</b> Peter Shen <peter.shen@biochem.utah.edu><br>
<b>Cc:</b> 3dem@ncmir.ucsd.edu<br>
<b>Subject:</b> Re: [3dem] weird ice?<o:p></o:p></span></p>
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<p class="MsoNormal"><o:p> </o:p></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">Hi Peter.<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">When I see this, I assumed I got my sample too dry, and what remains is protein film. I usually load a grid that should have “thicker ice”. The way I assess
which ones should have thicker ice is by recording both humidity in my vitrobot chamber and assessing blot size on my vitrobot filter paper immediately after the blot has happened (somewhat subjective, but with a single sample, side-entry cryoholder – I’ve
gotten pretty good at picking out which grids will have suitable ice). <o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">For example…(assuming everything else is the same)<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">lower humidity with a big blot on filter paper = thinner ice (maybe dried, as in your case)<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">lower humidity with smaller blot = thicker grid<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">higher humidity with small blot = thickest grid<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">My two cents, for what it’s worth!
</span><span style="font-size:11.0pt;font-family:Wingdings;color:#1F497D">J</span><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">HTH,<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">Tommi<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">Tommi A. White, Ph.D.<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">Assistant Professor of Biochemistry<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">Director, Electron Microscopy Core Facility<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">University of Missouri<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">W117 Veterinary Medicine Building<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">1600 East Rollins Street<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">Columbia, MO 65211<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">573-882-8304<o:p></o:p></span></p>
<p class="MsoNormal"><a href="mailto:WhiteTo@missouri.edu"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#0563C1">WhiteTo@missouri.edu</span></a><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p></o:p></span></p>
<p class="MsoNormal"><a href="http://emc.missouri.edu/"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#0563C1">http://emc.missouri.edu</span></a><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p> </o:p></span></p>
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<p class="MsoNormal"><b><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">From:</span></b><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"> 3dem [</span><a href="mailto:3dem-bounces@ncmir.ucsd.edu"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">mailto:3dem-bounces@ncmir.ucsd.edu</span></a><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">]
<b>On Behalf Of </b>Peter Shen<br>
<b>Sent:</b> Tuesday, October 16, 2018 6:39 PM<br>
<b>To:</b> </span><a href="mailto:3dem@ncmir.ucsd.edu"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">3dem@ncmir.ucsd.edu</span></a><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><br>
<b>Subject:</b> [3dem] weird ice?<o:p></o:p></span></p>
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<p class="MsoNormal"><o:p> </o:p></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma",sans-serif;color:black">Hi everyone,
<o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma",sans-serif;color:black"><o:p> </o:p></span></p>
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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:<o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma",sans-serif;color:black"><o:p> </o:p></span></p>
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<p class="MsoNormal"><a href="https://www.dropbox.com/sh/mlfado5rf9o447c/AAAt9VJM28KG6sCaNnCXIXQga?dl=0" target="_blank"><span style="font-size:10.0pt;font-family:"Tahoma",sans-serif">https://www.dropbox.com/sh/mlfado5rf9o447c/AAAt9VJM28KG6sCaNnCXIXQga?dl=0</span></a><span style="font-size:10.0pt;font-family:"Tahoma",sans-serif;color:black"><o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma",sans-serif;color:black"><o:p> </o:p></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... <o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma",sans-serif;color:black"><o:p> </o:p></span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma",sans-serif;color:black">Peter<o:p></o:p></span></p>
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