[3dem] [ext] Spots in micrographs

[Schacherl, Magdalena]

Hi Saumya

Did you check the same grids with another protein (e.g. apoferritin) in another buffer (but same blotting paper, same water source for the humidifier, etc.)?

We used to have the same problem very occasionally with our old Vitrobot Mark II (see attached micrograph). We think (but do not have the final proof) that it could have been a problem with the humidifier.


Good luck with future grids.

Best regards,
Magdalena


Von: 3dem <3dem-bounces at ncmir.ucsd.edu> im Auftrag von "Bajaj Saumya (Dr)" <Saumya.Bajaj at ntu.edu.sg>
Datum: Freitag, 20. März 2020 um 17:15
An: "3dem at ncmir.ucsd.edu" <3dem at ncmir.ucsd.edu>
Betreff: [ext] [3dem] Spots in micrographs

Dear all,

We are trying to solve the structure of a membrane protein. We have recently started seeing black spots (~1nm) on our micrographs (image attached). The pattern is not uniform, as there are areas where there are no spots. We tried changing buffer and detergent stock – that did not help. The grids are commercial graphene oxide girds and our buffer contains 0.03% GDN detergent.

Have any of you noticed something similar, or can help throw some light on what it could be?

Thanks a lot.
Best regards,
Saumya
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