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Sci Adv. 2018 Apr 20;4(4):eaaq1202. doi: 10.1126/sciadv.aaq1202. eCollection 2018 Apr.

The role of electron irradiation history in liquid cell transmission electron microscopy.

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Environmental Molecular Sciences Laboratory, 3335 Innovation Boulevard, Richland, WA 99354, USA.
Michigan Technological University, 1400 Townsend Drive, Houghton, MI 49931, USA.
Florida State University, 600 West College Avenue, Tallahassee, FL 32306, USA.
University of Illinois Chicago, 1200 West Harrison Street, Chicago, IL 60607, USA.
School of Biological Sciences, Washington State University, Pullman, WA 99164, USA.


In situ liquid cell transmission electron microscopy (LC-TEM) allows dynamic nanoscale characterization of systems in a hydrated state. Although powerful, this technique remains impaired by issues of repeatability that limit experimental fidelity and hinder the identification and control of some variables underlying observed dynamics. We detail new LC-TEM devices that improve experimental reproducibility by expanding available imaging area and providing a platform for investigating electron flux history on the sample. Irradiation history is an important factor influencing LC-TEM results that has, to this point, been largely qualitatively and not quantitatively described. We use these devices to highlight the role of cumulative electron flux history on samples from both nanoparticle growth and biological imaging experiments and demonstrate capture of time zero, low-dose images on beam-sensitive samples. In particular, the ability to capture pristine images of biological samples, where the acquired image is the first time that the cell experiences significant electron flux, allowed us to determine that nanoparticle movement compared to the cell membrane was a function of cell damage and therefore an artifact rather than visualizing cell dynamics in action. These results highlight just a subset of the new science that is accessible with LC-TEM through the new multiwindow devices with patterned focusing aides.

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