Inactivation of Fluorescent Lipid Bilayers by Irradiation With 300 keV Electrons Using Liquid Cell Transmission Electron Microscopy

Moser, Trevor; Evans, James E.

doi:10.3389/fnano.2022.772469

Title: Inactivation of Fluorescent Lipid Bilayers by Irradiation With 300 keV Electrons Using Liquid Cell Transmission Electron Microscopy

Journal Article · Tue Mar 29 00:00:00 EDT 2022 · Frontiers in Nanotechnology

DOI:https://doi.org/10.3389/fnano.2022.772469· OSTI ID:1860280

Moser, Trevor; Evans, James E.

Liquid cell transmission electron microscopy allows for imaging of samples in a fully hydrated state at high resolution and has the potential for visualizing static or dynamic biological structures. However, the ionizing nature of the electron beam makes it difficult to discern real physiological dynamics from radiation induced artifacts within liquid cell samples. Electron flux thresholds for achieving high resolution structures from biological samples frozen in ice have been described extensively by the cryo-electron microscopy field, while electron flux thresholds which do not result in a functional change for biological samples within the hydrated environment of a transmission electron microscope liquid cell is less clear. Establishing these functional thresholds for biologically relevant samples is important for accurate interpretation of results from liquid cell experiments. Here we demonstrate the electron damage threshold of fluorescently tagged lipid bilayers by quantifying the change in fluorescence before and after electron exposure. We observe the reduction of fluorescent signal in bilayers by 25% after only 0.0005 e ⁻ /Å ² and a reduction of over 90% after 0.01 e ⁻ /Å ² . These results indicate that the loss of function occurs at irradiation thresholds far below a typical single high resolution (scanning) transmission electron microscopy image and orders of magnitude below fluxes used for preserving structural features with cryo-electron microscopy.

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Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

OSTI ID:: 1860280

Journal Information:: Frontiers in Nanotechnology, Journal Name: Frontiers in Nanotechnology Vol. 4; ISSN 2673-3013

Publisher:: Frontiers Media SACopyright Statement

Country of Publication:: Switzerland

Language:: English

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