[3dem] C-flat grids cryo-EM with manual blotting
Jean Watermeyer
jeanwatermeyer at gmail.com
Wed Dec 10 01:29:04 PST 2008
Hi all
Is there anyone on this list who has been doing cryo-EM with C-flat grids,
using manual blotting, and glow-discharging?
We have been trying to optimise our blotting for the thinnest possible ice
and have found several problems:
- if the blotting is too rough or the plunging too deep, all of the carbon
comes off the grid
- if we glow-discharge for 10s at 20 mA, blotting for ~8 s at 50% humidity,
we get ice that is much too thick
- without glow-discharging, we get too steep a gradient of ice thickness
We'd really appreciate hearing from anyone who has managed to get very thin
ice with these grids using manual blotting.
Sincerely
Jean Watermeyer
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