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<pre><span style="font-family:"Calibri","sans-serif";color:#1F497D">Hi Nadav:<o:p></o:p></span></pre>
<pre><span style="font-family:"Calibri","sans-serif";color:#1F497D"><o:p> </o:p></span></pre>
<pre><span style="font-family:"Calibri","sans-serif";color:#1F497D">I have similar observation like Garry that quantifoil grids gave such problem but C-flat is fine, using the same preparation conditions. Originally I thought it was the ethane contamination. I try to shoot the same position two or three times. The contaminations got fade. The protein particles were fine. Did you check the sample right after the grid preparation?<o:p></o:p></span></pre>
<pre><span style="font-family:"Calibri","sans-serif";color:#1F497D"><o:p> </o:p></span></pre>
<pre><span style="font-family:"Calibri","sans-serif";color:#1F497D">Xiaodi<o:p></o:p></span></pre>
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<p class="MsoNormal"><b><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif"">From:</span></b><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif""> 3dem [mailto:3dem-bounces@ncmir.ucsd.edu]
<b>On Behalf Of </b>Eric Gilles Hanssen<br>
<b>Sent:</b> Thursday, September 1, 2016 3:02 PM<br>
<b>To:</b> Nadav Elad<br>
<b>Cc:</b> 3dem@ncmir.ucsd.edu<br>
<b>Subject:</b> Re: [3dem] contamination in cryo samples<o:p></o:p></span></p>
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<p class="MsoNormal">Hi Nadav,<o:p></o:p></p>
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<p class="MsoNormal">We had exactly the same thing happening in our lab. The humidity in the room was way to high after some renovations we undertook. We dropped the room humidity to ~30% which made things much better, we still had some slight issues but after
changing the zeolite (that was believe or not 10 years old) in our 626 for the new carbon pack the issue completely disappeared.<o:p></o:p></p>
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<p class="MsoNormal">Regards<o:p></o:p></p>
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<p class="MsoNormal">Eric<o:p></o:p></p>
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<p class="MsoNormal"><o:p> </o:p></p>
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<p class="MsoNormal"><span style="font-size:13.0pt">----------------------------------------------------------------------------</span><o:p></o:p></p>
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<p class="MsoNormal">A/Prof Eric Hanssen<o:p></o:p></p>
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<p class="MsoNormal"><o:p> </o:p></p>
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<p class="MsoNormal">Head - Advanced Microscopy Facility<o:p></o:p></p>
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<p class="MsoNormal">Adjunct Associate Professor - Department of Biochemistry and Molecular Biology<o:p></o:p></p>
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<p class="MsoNormal"><br>
<br>
<o:p></o:p></p>
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<p class="MsoNormal">Bio21 Molecular Science and Biotechnology Institute<o:p></o:p></p>
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<p class="MsoNormal">30 Flemington Road - The University of Melbourne - Victoria 3010 - Australia<o:p></o:p></p>
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<p class="MsoNormal">email: <a href="mailto:ehanssen@unimelb.edu.au">ehanssen@unimelb.edu.au</a> | Office: +61 3 83442449 | Microscope: +61 3 83442509 | Fax: +61 3 9347 826<o:p></o:p></p>
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<p class="MsoNormal"><o:p> </o:p></p>
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On 1 Sep 2016, at 02:01, Nadav Elad <<a href="mailto:nadav.elad@weizmann.ac.il">nadav.elad@weizmann.ac.il</a>> wrote:<o:p></o:p></p>
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<p class="MsoNormal">Dear all<o:p></o:p></p>
<p class="MsoNormal"> <o:p></o:p></p>
<p class="MsoNormal">We are having a serious contamination problem in our cryo preparations and would be grateful for any suggestion. Although this issue has been discussed in the past in this forum, the solutions raised do not seem to work. The attached image
shows the contamination on a bad day. I am referring mainly to the lighter, smaller particles (10-20 nm) which are the most persistent. They seem to be above or below the ice layer and are the first to evaporate under the electron beam. The concentration of
contaminants is variable. <o:p></o:p></p>
<p class="MsoNormal"> <o:p></o:p></p>
<p class="MsoNormal">We are using a Leica EM GP plunger for freezing, and Tecnai T12 and F20 microscopes with Gatan side entry holders. We have tried to replace the ethane cylinder and are in the process of ordering a third one, cleaning the glow discharger
machines, different cryo holders, different cryo-transferring units and grids of varying sources. There seems to be no leaks in the compustages of the microscopes.<o:p></o:p></p>
<p class="MsoNormal"> <o:p></o:p></p>
<p class="MsoNormal">Thank you for your help<o:p></o:p></p>
<p class="MsoNormal"> <o:p></o:p></p>
<p class="MsoNormal">Nadav<o:p></o:p></p>
<p class="MsoNormal"> <o:p></o:p></p>
<p class="MsoNormal">--<o:p></o:p></p>
<p class="MsoNormal">Nadav Elad, Ph.D<span dir="RTL"></span><span lang="HE" dir="RTL" style="font-family:"Arial","sans-serif""><span dir="RTL"></span>.<br>
</span>Electron Microscopy Unit<span lang="HE" dir="RTL" style="font-family:"Arial","sans-serif""><br>
</span>Weizmann Institute of Science<o:p></o:p></p>
<p class="MsoNormal">Rehovot, Israel<span lang="HE" dir="RTL" style="font-family:"Arial","sans-serif""><br>
</span>Tel: +972 (0) 8 934 2115<o:p></o:p></p>
<p class="MsoNormal">Mobile: +972 (0) 52 3909 420<o:p></o:p></p>
<p class="MsoNormal">Email: <a href="mailto:nadav.elad@weizmann.ac.il">nadav.elad@weizmann.ac.il</a><o:p></o:p></p>
<p class="MsoNormal"> <o:p></o:p></p>
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<p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Times New Roman","serif""><contamination.jpg><o:p></o:p></span></p>
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