[3dem] C-flat grids cryo-EM with manual blotting

[Jean Watermeyer]

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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