[3dem] C-flats

Wed May 25 13:10:24 PDT 2011

Hi Thorsten,

same here with CF-2/4 and Gatan Cp3 plunger. It worked fine ONLY when skipping the glow-discharge. Even a gentle treatment with the Gatan plasma cleaner, for only 5-10 seconds, would break the C-foil completely.

I had Protochips replace all remaining sealed boxes (more than 30) with a slightly thicker carbon, and had no problems since.  Protochips people were very fast in their turn around and replaced all the boxes at no additional cost.

Best of luck,
Sacha

=============================
Sacha De Carlo

________________________________
From: Thorsten Mielke <mielke at molgen.mpg.de>
To: 3dem at ncmir.ucsd.edu
Sent: Wednesday, May 25, 2011 11:08 AM
Subject: [3dem] C-flats

Dear all,

we are trying to vitrify some small proteins on c-flat grids (type CF-2/4-4C) using a Vitrobot from FEI. However, after plunge-freezing, the carbon film is heavily destroyed. Additionally, we find only a few good holes on the remaining intact meshes. The carbon film also seems to be extremely fragile and easily disrupts upon focusing (300 kV, 155kx Polara). The grids seem to be OK (with or without glow-discharching) until plunge-freezing. We varied the blotting offset at the Vitrobot to play with the blotting force, no success.  Did you observe similar problems with C-flats? Any idea how to prevent the carbon film from breaking?

Thanks

Thorsten

-- 
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Dr. Thorsten Mielke
- UltraStrukturNetzwerk -

Max Planck Institute for Molecular Genetics        
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Tel.: +49-(0)30 8413-1644/1141
Fax: +49-(0)30 8413-1385
E-mail: mielke at molgen.mpg.de

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