[3dem] Insertion of Holder 626

[Schwartz, Cindi (NIH/NIAID) [E]]

In the past with our 626 holders, I had to replace the zeolite every once in a while--maybe 3 times in 10+ years??? Ours were charcoal at the beginning and then we switched to zeolite. 

At any rate, you can have Gatan do it, or they talked me through it and it's a very easy procedure and helped immediately.

Cheers,
Cindi L. Schwartz 

Electron Microscopist
Rocky Mountain Labs/NIAID/NIH
903 South 4th Street
Hamilton, MT  59840
406-363-9228
Cindi.Schwartz at NIH.gov
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-----Original Message-----
From: TALAVERA PEREZ Ariel <Ariel.Talavera at ulb.be> 
Sent: Monday, February 24, 2020 6:13 AM
To: 3dem at ncmir.ucsd.edu
Subject: [3dem] Insertion of Holder 626

Dear all,

I am using a Gatan 626 holder on a side entry TALOS microscope. I am getting problems with very frequent crystalline ice formation on the grids. Some times I get a mix of vitreous and crystalline water, but must of the time I only get crystalline water. I have been extra careful during sample preparation, and transferring the grind on to the holder, and I have done 48 hours holder bake out before insertion, but I still get ice on the grid.

I monitored the temperature of the holder during the insertion process, including 3 minutes for the Airlock pump. During those 3 minutes the temperature rises up to -152.2. Can this be the reason of the unwanted ice formation? How high can be the temperature of the holder to ensure a nice vitreous water?

I am also getting vacuum problems while inserting the holder. This process has resulted to be extremely, extremely sensitive. After the 3 minutes Airlock pumping (I cannot give longer time because of the aforementioned temperature issue) as soon as I open the column port the vacuum either rises to 99 Log or completely crashes the column vacuum. 
This has happened with two different holders either cooled with liquid nitrogen or at room temperature. So, my question: Is this insertion process always that  picky or it can be that there is something wrong with the holders/Airlock pump?

Thanks a lot in advanced.

Best regards.

Ariel

--
Ariel TALAVERA PEREZ, PhD
Center for Microscopy and Molecular Imaging (CMMI) Laboratory of Microscopy Université Libre de Bruxelles Rue Adrienne Bolland, 8
B-6041 Gosselies, Belgium
Phone +32 (0)26509866

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