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<p class="MsoNormal"><span style="font-size:11.0pt; font-family:"Calibri",sans-serif; color:#1F497D">Hi
<span class="SpellE">everyone,</span></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt; font-family:"Calibri",sans-serif; color:#1F497D"> </span></p>
<p class="MsoNormal"><span style="font-size:11.0pt; font-family:"Calibri",sans-serif; color:#1F497D">My original tests for the PNAS paper in 2014 were performed on an F20. There, we observed a correlation between the vacuum level (using or not the anticontamination
device) and the recovery speed and concluded that the effect is due to the interaction of the
</span><span style="font-size:11.0pt; font-family:"Calibri",sans-serif; color:#1F497D">beam-modified surface of the amorphous carbon with the
</span><span style="font-size:11.0pt; font-family:"Calibri",sans-serif; color:#1F497D">residual gasses or the atomic species and radicals created by the ion pump. On the F20 the recovery took a few days. The vacuum in a Titan
<span class="SpellE">Krios</span> is much better than in an F20 and my subjective experience is that the spots take much longer to recover there. I’ve never had the chance to test the recovery speed on our
<span class="SpellE">Krios</span> because the phase plate is used a lot and we want to spend the limited time we have on the <span class="SpellE"> Krios</span> for actual data acquisition.<br>
</span></p>
<p class="MsoNormal"><span style="font-size:11.0pt; font-family:"Calibri",sans-serif; color:#1F497D">After 2 1/2 years of use our Krios phase plate finally reached a point where we had to change it. There were too many dust-like contaminants on the surface
which limited the usable areas to less than one third of the original areas. Those contaminants get deposited either during venting of the column for service or if one forgets to retract the phase plate when loading/unloading samples. Also, the non-contaminated
areas were showing too much astigmatism variation from spot to spot. This could have been because of the "scars" that Matthias described.</span></p>
<p class="MsoNormal"><span style="font-size:11.0pt; font-family:"Calibri",sans-serif; color:#1F497D"><br>
</span></p>
<p class="MsoNormal"><span style="font-size:11.0pt; font-family:"Calibri",sans-serif; color:#1F497D">Cheers,</span></p>
<p class="MsoNormal"><span style="font-size:11.0pt; font-family:"Calibri",sans-serif; color:#1F497D"><br>
</span></p>
<p class="MsoNormal"><span style="font-size:11.0pt; font-family:"Calibri",sans-serif; color:#1F497D">Rado</span></p>
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</span></p>
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</span></p>
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<p class="MsoNormal"><a name="_MailOriginal"><b><span style="font-size:11.0pt; font-family:"Calibri",sans-serif">From:</span></b></a><span style=""><span style="font-size:11.0pt; font-family:"Calibri",sans-serif"> 3dem [mailto:3dem-bounces@ncmir.ucsd.edu]
<b>On Behalf Of </b>Craig Yoshioka<br>
<b>Sent:</b> Friday, March 24, 2017 19:28<br>
<b>To:</b> Anchi Cheng <acheng@nysbc.org><br>
<b>Cc:</b> 3dem@ncmir.ucsd.edu<br>
<b>Subject:</b> Re: [3dem] volta phase plate recovery time</span></span></p>
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<p class="MsoNormal"><span style=""><span style="">Hi Anchi, </span></span></p>
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<p class="MsoNormal"><span style=""><span style=""> </span></span></p>
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<p class="MsoNormal"><span style=""><span style="">By the time we get through all the spots on all our films it usually has been over 2 months… I’m not 100% sure if all the old spots are gone… but I have not seen them, nor have I noticed their effects on the
images after doing the full cycle. When I have worried about it before in the past, I’ve just offset all the positions by using the motorized aperture controls to shift the whole phase plate. </span></span></p>
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<p class="MsoNormal"><span style=""><span style="">Rado can correct me if I’m wrong, but I think secondary spots have the most deleterious effect if fairly close where they affect a large amount of low-resolution scattering. I think if they are far enough
out you would probably not notice their effect, and it probably would not be a showstopper anyways with random particle orientations and averaging. </span></span></p>
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<p class="MsoNormal"><span style=""><span style="">On Mar 24, 2017, at 11:20 AM, Anchi Cheng <</span></span><a href="mailto:acheng@nysbc.org" target="_blank"><span style=""><span style="">acheng@nysbc.org</span></span><span style=""></span></a><span style=""><span style="">>
wrote:</span></span></p>
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<p class="MsoNormal"><span style=""><span style=""> </span></span></p>
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<p class="MsoNormal"><span style=""><span style="">Hi, Wim, </span></span></p>
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<p class="MsoNormal"><span style=""><span style=""> </span></span></p>
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<p class="MsoNormal"><span style=""><span style="">Thanks for replying. No, these are regular-spaced spots that show up after</span></span></p>
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<p class="MsoNormal"><span style=""><span style="">using the patch position. Similar to Figure 2 in Rado’s 2015 PNAS paper.</span></span></p>
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<p class="MsoNormal"><span style=""><span style="">The spacing corresponds also the distance between the defined distance</span></span></p>
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<p class="MsoNormal"><span style=""><span style="">between patches. Incidentally, this phase plate is the cleanest I have ever</span></span></p>
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<p class="MsoNormal"><span style=""><span style="">seen. It was hard to find a feature on it to align.</span></span></p>
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<p class="MsoNormal"><span style=""><span style=""> </span></span></p>
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<p class="MsoNormal"><span style=""><span style="">We do have some burn spots disappeared after 1.5 month. There were the</span></span></p>
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<p class="MsoNormal"><span style=""><span style="">ones that was irradiated less.</span></span></p>
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<p class="MsoNormal"><span style=""><span style=""> </span></span></p>
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<p class="MsoNormal"><span style=""><span style="">I am hoping I get a number from you or others with phase plates longer than</span></span></p>
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<p class="MsoNormal"><span style=""><span style="">we have how long you wait before using the same phase plate patch position</span></span></p>
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<p class="MsoNormal"><span style=""><span style="">again.</span></span></p>
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<p class="MsoNormal"><span style=""><span style=""> </span></span></p>
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<p class="MsoNormal"><span style=""><span style="">Best,</span></span></p>
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<p class="MsoNormal"><span style=""><span style=""> </span></span></p>
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<p class="MsoNormal"><span style=""><span style="">Anchi</span></span></p>
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<p class="MsoNormal"><span style=""><span style=""> </span></span></p>
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<p class="MsoNormal"><span style=""><span style="">On Mar 24, 2017, at 4:15 AM, Wim Hagen <</span></span><a href="mailto:wim.hagen@embl.de" target="_blank"><span style=""><span style="">wim.hagen@embl.de</span></span><span style=""></span></a><span style=""><span style="">>
wrote:</span></span></p>
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<p class="MsoNormal"><span style=""><span style=""> </span></span></p>
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<p class="MsoNormal"><span style=""><span style="font-family:"Tahoma",sans-serif">Hi Anchi,<br>
<br>
Maybe it is contamination? I see some spots in some areas on our brand new phase plate, the contrast of the spots is reversed w.r.g. to a Volta patch. Here are some images with many bad spots and one 'quick and dirty fresh' Volta patch, 'above and below' plane:<br>
<br>
</span></span><a href="https://oc.embl.de/index.php/s/j3SlNyNwsspH00U" target="_blank"><span style=""><span style="font-family:"Tahoma",sans-serif">https://oc.embl.de/index.php/s/j3SlNyNwsspH00U</span></span><span style=""></span></a><span style=""><span style="font-family:"Tahoma",sans-serif"><br>
<br>
Best,<br>
<br>
Wim</span></span></p>
<p class="MsoNormal"><span style=""><span style="font-family:"Tahoma",sans-serif"> </span></span></p>
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<p class="MsoNormal"><span style=""><span style="font-family:"Tahoma",sans-serif">On Thu, Mar 23, 2017 at 18:33, Anchi Cheng <</span></span><a href="mailto:acheng@nysbc.org" target="_blank"><span style=""><span style="font-family:"Tahoma",sans-serif">acheng@nysbc.org</span></span><span style=""></span></a><span style=""><span style="font-family:"Tahoma",sans-serif">>
wrote:</span></span></p>
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<blockquote style="margin-top:5.0pt; margin-bottom:5.0pt">
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<p class="MsoNormal"><span style=""><span style="font-family:"Tahoma",sans-serif">Hi, all,<br>
<br>
I am wondering what people’s experience in the length of time<br>
they wait before they reuse a patch position on FEI volta phase plate.<br>
<br>
We can still see rows of the different contrast (burn spot, we call it) at low mag<br>
and very high defocus (-100 mm) after a month and a half.<br>
<br>
If I return to a patch in 2 weeks, the phase shift development is faster. In addition,<br>
there is no guarantee that I am back at the exact position.<br>
<br>
Anchi<br>
<br>
NRAMM<br>
Simons Microscopy Microscopy Center<br>
New York Structural Biology Center<br>
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