[3dem] C-flats
Thorsten Mielke
mielke at molgen.mpg.de
Wed May 25 02:08:32 PDT 2011
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
--
*******************************************
Dr. Thorsten Mielke
- UltraStrukturNetzwerk -
Max Planck Institute for Molecular Genetics
Ihnestr. 63-73
D-14195 Berlin
Tel.: +49-(0)30 8413-1644/1141
Fax: +49-(0)30 8413-1385
E-mail: mielke at molgen.mpg.de
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