[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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