Microscale Fluid Behavior during Cryo-EM Sample Blotting.

ABSTRACT: Blotting has been the standard technique for preparing aqueous samples for single-particle electron cryo-microscopy for over three decades. This technique removes the excess solution from a transmission electron microscope grid by pressing absorbent filter paper against the specimen before vitrification. However, this standard technique produces vitreous ice with inconsistent thickness from specimen to specimen and from region to region within the same specimen, the reasons for which are not understood. Here, high-speed interference contrast microscopy is used to demonstrate that the irregular pattern of fibers in the filter paper imposes tortuous, highly variable boundaries during the removal of excess liquid from a flat, hydrophilic surface. As a result, aqueous films of nonuniform thick

SUBMITTER: Armstrong M 

PROVIDER: S-EPMC7004840 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature