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<p class="MsoNormal"><span style="color:#1F497D">Hi Saumya, <o:p></o:p></span></p>
<p class="MsoNormal"><span style="color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="color:#1F497D">We have had that discussion a while back, we also discussed this at the Keystone cryoEM a couple of years ago. It seems you are not the only one having that issue. We definitely encountered it a fair few times.
However my gut feeling at the time was that the culprit was our glow discharge system. But I only had exclusion proofs (not the tweezers, not other proteins, not rust on the grids ….).
<o:p></o:p></span></p>
<p class="MsoNormal"><span style="color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="color:#1F497D">Since then we purchased a new glow discharge system. I have not seen these spots since. Saying that I am a little less on the microscope than 2 years ago but if some of my staffs or users are reading this please
feel free to contradict me. <o:p></o:p></span></p>
<p class="MsoNormal"><span style="color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="color:#1F497D">Maybe give it a good clean and try again.
<o:p></o:p></span></p>
<p class="MsoNormal"><span style="color:#1F497D">Cheers<o:p></o:p></span></p>
<p class="MsoNormal"><span style="color:#1F497D">Eric<o:p></o:p></span></p>
<p class="MsoNormal"><span style="color:#1F497D">(PS I am quite sure I will see them again now I have said that …)<o:p></o:p></span></p>
<p class="MsoNormal"><span style="color:#1F497D"><o:p> </o:p></span></p>
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<p class="MsoNormal"><b><span style="font-family:"Comic Sans MS";color:navy;mso-fareast-language:EN-AU">-----------------</span></b><b><span style="font-size:10.0pt;font-family:"Comic Sans MS";color:navy;mso-fareast-language:EN-AU"><o:p></o:p></span></b></p>
<p class="MsoNormal"><b><span style="font-family:"Comic Sans MS";color:navy;mso-fareast-language:EN-AU">Assoc. Prof Eric Hanssen<o:p></o:p></span></b></p>
<p class="MsoNormal"><b><span style="font-family:"Comic Sans MS";color:navy;mso-fareast-language:EN-AU"><o:p> </o:p></span></b></p>
<p class="MsoNormal"><b><span style="font-family:"Comic Sans MS";color:#1F497D;mso-fareast-language:EN-AU">Head - Advanced Microscopy Facility
<o:p></o:p></span></b></p>
<p class="MsoNormal"><b><span style="font-family:"Comic Sans MS";color:#1F497D;mso-fareast-language:EN-AU">Honorary Principal Fellow – Department of Biochemistry and Molecular Biology<o:p></o:p></span></b></p>
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<p class="MsoNormal"><b><span style="font-family:"Comic Sans MS";color:#1F497D;mso-fareast-language:EN-AU">President Australian Microscopy and Microanalysis Society<o:p></o:p></span></b></p>
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Bio21 Molecular Science and Biotechnology Institute<br>
</span><span style="font-family:"Comic Sans MS";color:gray;mso-fareast-language:EN-AU">30 Flemington Road - The University of Melbourne - Victoria 3010 - Australia<br>
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<a href="http://www.microscopy.unimelb.edu.au/"><span style="color:blue">www.microscopy.unimelb.edu.au</span></a><o:p></o:p></span></p>
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<p class="MsoNormal"><b><span lang="EN-US" style="mso-fareast-language:EN-AU">From:</span></b><span lang="EN-US" style="mso-fareast-language:EN-AU"> 3dem [mailto:3dem-bounces@ncmir.ucsd.edu]
<b>On Behalf Of </b>Bajaj Saumya (Dr)<br>
<b>Sent:</b> Friday, 20 March 2020 8:50 PM<br>
<b>To:</b> 3dem@ncmir.ucsd.edu<br>
<b>Subject:</b> [3dem] Spots in micrographs<o:p></o:p></span></p>
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<p class="MsoNormal"><o:p> </o:p></p>
<p class="MsoNormal"><span lang="EN-SG">Dear all,<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-SG"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-SG">We are trying to solve the structure of a membrane protein. We have recently started seeing black spots (~1nm) on our micrographs (image attached). The pattern is not uniform, as there are areas where there are no spots.
We tried changing buffer and detergent stock – that did not help. The grids are commercial graphene oxide girds and our buffer contains 0.03% GDN detergent.
<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-SG"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-SG">Have any of you noticed something similar, or can help throw some light on what it could be?
<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-SG"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-SG">Thanks a lot.<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-SG">Best regards,<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-SG">Saumya<o:p></o:p></span></p>
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