9. Immediately after blotting place the grid (carbon side down)
on top of a drop of 200 μL of a 2% (w/v) uranyl acetate
solution for 40 s. Transfer the grid to a second drop of
200 μL of a 2% (w/v) uranyl acetate solution and stain for
another 40 sec without blotting in between the two staining
steps. After staining, the grid is blotted from the back with filter
paper in order to remove the staining solution and dried on air
for at least 20 min prior to further use (see Note 9).
10. Inspect the nature of the granular aggregates and lattice quality
of the (fragmented) microcrystals by collecting TEM images
(see Note 10 and Fig. 4).
Fig. 3 Schematic representation of the blotting process
Detection of Microcrystals for CryoEM
303
on top of a drop of 200 μL of a 2% (w/v) uranyl acetate
solution for 40 s. Transfer the grid to a second drop of
200 μL of a 2% (w/v) uranyl acetate solution and stain for
another 40 sec without blotting in between the two staining
steps. After staining, the grid is blotted from the back with filter
paper in order to remove the staining solution and dried on air
for at least 20 min prior to further use (see Note 9).
10. Inspect the nature of the granular aggregates and lattice quality
of the (fragmented) microcrystals by collecting TEM images
(see Note 10 and Fig. 4).
Fig. 3 Schematic representation of the blotting process
Detection of Microcrystals for CryoEM
303