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<p class="MsoNormal"><span lang="EN-US">Hi Saumya<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">Did you check the same grids with another protein (e.g. apoferritin) in another buffer (but same blotting paper, same water source for the humidifier, etc.)?
<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">We used to have the same problem very occasionally with our old Vitrobot Mark II (see attached micrograph). We think (but do not have the final proof) that it could have been a problem with the humidifier.<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">Good luck with future grids.<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">Best regards,<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">Magdalena<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-US"><o:p> </o:p></span></p>
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<p class="MsoNormal"><b><span style="font-size:12.0pt;color:black">Von: </span></b><span style="font-size:12.0pt;color:black">3dem <3dem-bounces@ncmir.ucsd.edu> im Auftrag von "Bajaj Saumya (Dr)" <Saumya.Bajaj@ntu.edu.sg><br>
<b>Datum: </b>Freitag, 20. März 2020 um 17:15<br>
<b>An: </b>"3dem@ncmir.ucsd.edu" <3dem@ncmir.ucsd.edu><br>
<b>Betreff: </b>[ext] [3dem] Spots in micrographs</span><span style="font-size:12.0pt;color:black;mso-fareast-language:DE"><o:p></o:p></span></p>
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<p class="MsoNormal"><span lang="EN-US">Dear all,<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US"> <o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">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-US"> <o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">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-US"> <o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">Thanks a lot.<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">Best regards,<o:p></o:p></span></p>
<p class="MsoNormal">Saumya</p>
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