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Hi Ariel
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<div class="">To add to Krishna’s comments the airlock my also need a clean. I recently had frequent vacuum crashes on our CM200 and a careful clean of the airlock tube removed a lot of particulates that were the cause of the problem. </div>
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<div class="">I would recommend you ask you field service engineer to clean it as there are some sensitive components that you don't want to damage.</div>
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<div class="">Best wishes</div>
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<div class="">Christos</div>
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Dr. Christos Savva</div>
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Facility Manager</div>
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Midlands Regional Cryo-EM Facility</div>
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University of Leicester</div>
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Henry Wellcome Building</div>
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Lancaster Road</div>
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Leicester, LE1 7HB</div>
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<b class=""><span class="" style="font-size: 10pt; color: rgb(187, 0, 0);">t:</span></b><span class="" style="font-size: 10pt;"> +44 116 229 7109<br class="">
</span><b class=""><span class="" style="font-size: 10pt; color: rgb(187, 0, 0);">e:</span></b><span class="" style="font-size: 10pt;"> </span><span class="" style="font-size: 10pt; font-family: "Times New Roman", serif;"><a href="mailto:gwcs1@le.ac.uk" class="" target="_blank" rel="noopener noreferrer"><span class="" style="font-family: Helvetica; color: blue;">gwcs1@le.ac.uk</span></a></span><span class="" style="font-size: 10pt;"><br class="">
</span><b class=""><span class="" style="font-size: 10pt; color: rgb(187, 0, 0);">w:</span></b><u class=""><span class="" style="font-size: 10pt; color: blue;"><a href="http://www.le.ac.uk/cryoem" class="" target="_blank" rel="noopener noreferrer"><span class="" style="color: blue;">www.le.ac.uk/cryoem</span></a></span></u></div>
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<blockquote type="cite" class="">
<div class="">On 24 Feb 2020, at 13:13, TALAVERA PEREZ Ariel <<a href="mailto:Ariel.Talavera@ulb.be" class="">Ariel.Talavera@ulb.be</a>> wrote:</div>
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<div class="">Dear all,<br class="">
<br class="">
I am using a Gatan 626 holder on a side entry TALOS microscope. I am <br class="">
getting problems with very frequent crystalline ice formation on the <br class="">
grids. Some times I get a mix of vitreous and crystalline water, but <br class="">
must of the time I only get crystalline water. I have been extra careful <br class="">
during sample preparation, and transferring the grind on to the holder, <br class="">
and I have done 48 hours holder bake out before insertion, but I still <br class="">
get ice on the grid.<br class="">
<br class="">
I monitored the temperature of the holder during the insertion process, <br class="">
including 3 minutes for the Airlock pump. During those 3 minutes the <br class="">
temperature rises up to -152.2. Can this be the reason of the unwanted <br class="">
ice formation? How high can be the temperature of the holder to ensure a <br class="">
nice vitreous water?<br class="">
<br class="">
I am also getting vacuum problems while inserting the holder. This <br class="">
process has resulted to be extremely, extremely sensitive. After the 3 <br class="">
minutes Airlock pumping (I cannot give longer time because of the <br class="">
aforementioned temperature issue) as soon as I open the column port the <br class="">
vacuum either rises to 99 Log or completely crashes the column vacuum. <br class="">
This has happened with two different holders either cooled with liquid <br class="">
nitrogen or at room temperature. So, my question: Is this insertion <br class="">
process always that picky or it can be that there is something wrong <br class="">
with the holders/Airlock pump?<br class="">
<br class="">
Thanks a lot in advanced.<br class="">
<br class="">
Best regards.<br class="">
<br class="">
Ariel<br class="">
<br class="">
-- <br class="">
Ariel TALAVERA PEREZ, PhD<br class="">
Center for Microscopy and Molecular Imaging (CMMI)<br class="">
Laboratory of Microscopy<br class="">
Université Libre de Bruxelles<br class="">
Rue Adrienne Bolland, 8<br class="">
B-6041 Gosselies, Belgium<br class="">
Phone +32 (0)26509866<br class="">
<br class="">
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