[3dem] Negative stain on graphene oxyde grids

[Matthews-Palmer, Teige Rowan Seal]

Dear Julien,
It’s a good idea, to check that your sample will adsorb to GrOx, and perhaps you’d also like to see how it’s oriented.
However, things might look different between a negative stain protocol versus blotting and plunge freezing. It’s probably better to move quickly to freezing, as that’s where optimisation can take time, and you can see how your sample interacts with GrOx under the conditions you’ll use for eventual data collection. In other words, diagnose GrOx at stage 3 of this scheme: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5140023/figure/F1/
If neg stain on GrOx turns out to be challenging, it will be a waste of time for likely no benefit over regular grids.
Negative stain on regular carbon grids can tell you if your particle looks the right shape & size, and what the concentration is like. (A rule of thumb some use to start, is having found a good concentration in neg stain, 10x for cryo in holes, and a smaller increase for cryo on support films.)

All the best
Teige
On 11 Sep 2018, at 16:55, Julien Bous <julien.bous at etu.umontpellier.fr<mailto:julien.bous at etu.umontpellier.fr>> wrote:


Dear Community,

I'm PhD student in cryoEM and I plan to use graphene oxide grids for a complicate sample. Therefore, in order to make a first assessment of the interaction between the graphene oxide and my sample, I want to try in first to do a negative stain on graphene oxide with my sample Has anyone ever seen or tried this , I didn't find any bibliography? And is it looks like a good idea for more experienced peoples?

Thank you very much for your time.

Best,

Julien

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