[3dem] protein can't go to holes of cryo EM grids

[Rajiv Ranjan Singh]

Hey Sasha,

Try H2, O2 Plasma Cleaning for 30S in each case. If you see good particles
in Negative stains.

I hope you are doing plasma cleaning before coating GO, if not then try
following protocol from Eva Nogales lab.

Best,
Rajiv Ranjan Singh



On Tue, Oct 11, 2022 at 4:47 AM SSS Sasha <sss_jiandan at outlook.com> wrote:

> Dear all,
>
> I was trying to freeze cryo EM grids for a protein ~ 0.73MDa under several
> conditions, but it didn't work well. I make a brief summary here. Does
> anyone have any suggestions? Thanks in advance!
>
> Holly grids CF-1.2/1.3-4Cu-50: plasma cleaning for 10s using O2,Ar2
>     Protein can't go in to the hole
>
> Lacey carbon 400mesh Cu:  plasma cleaning for 15s using O2, Ar2
> Contrast is quite low, hardly see the particles
>
> GO on Quantifoil: plasma cleaning for 15s using H2, O2
> Protein can't go into hole
>
> Best regards,
> Shasha
>
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