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Immobilized particles can still be contacted by the air-water interface if the remaining buffer is too thin. Although use of affinity support films may provide a path to reliably prepare cryo-grids for every type of specimen, a remaining problem is to find a way to keep the air-water interface from touching the immobilized particles. As the green-colored interface indicates, the situation currently is safe, but further thinning, as suggested by the arrows, may not be a good thing.
