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3D printed cell culture grid holders for improved cellular specimen preparation in cryo-electron microscopy

View ORCID ProfileFlorian Fäßler, Bettina Zens, View ORCID ProfileRobert Hauschild, View ORCID ProfileFlorian K.M. Schur
doi: https://doi.org/10.1101/2020.06.12.147678
Florian Fäßler
1Institute of Science and Technology Austria, Am Campus 1, Klosterneuburg, Austria
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Bettina Zens
1Institute of Science and Technology Austria, Am Campus 1, Klosterneuburg, Austria
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Robert Hauschild
1Institute of Science and Technology Austria, Am Campus 1, Klosterneuburg, Austria
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Florian K.M. Schur
1Institute of Science and Technology Austria, Am Campus 1, Klosterneuburg, Austria
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  • For correspondence: florian.schur@ist.ac.at
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Abstract

Cryo-electron microscopy (cryo-EM) of cellular specimens provides insights into biological processes and structures within a native context. However, a major challenge still lies in the efficient and reproducible preparation of adherent cells for subsequent cryo-EM analysis. This is due to the sensitivity of many cellular specimens to the varying seeding and culturing conditions required for EM experiments, the often limited amount of cellular material and also the fragility of EM grids and their substrate. Here, we present low-cost and reusable 3D printed grid holders, designed to improve specimen preparation when culturing challenging cellular samples directly on grids. The described grid holders increase cell culture reproducibility and throughput, and reduce the resources required for cell culturing. We show that grid holders can be integrated into various cryo-EM workflows, including micro-patterning approaches to control cell seeding on grids, and for generating samples for cryo-focused ion beam milling and cryo-electron tomography experiments. Their adaptable design allows for the generation of specialized grid holders customized to a large variety of applications.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted June 12, 2020.
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3D printed cell culture grid holders for improved cellular specimen preparation in cryo-electron microscopy
Florian Fäßler, Bettina Zens, Robert Hauschild, Florian K.M. Schur
bioRxiv 2020.06.12.147678; doi: https://doi.org/10.1101/2020.06.12.147678
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3D printed cell culture grid holders for improved cellular specimen preparation in cryo-electron microscopy
Florian Fäßler, Bettina Zens, Robert Hauschild, Florian K.M. Schur
bioRxiv 2020.06.12.147678; doi: https://doi.org/10.1101/2020.06.12.147678

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