期刊
JOURNAL OF STRUCTURAL BIOLOGY
卷 212, 期 3, 页码 -出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.jsb.2020.107633
关键词
Electron microscopy; Cryo-EM; EM sample preparation; 3D printing; Cell culture
资金
- Austrian Science Fund (FWF) [P33367]
- Niederosterreich Fond
- Scientific Service Units (SSU) of IST Austria by Scientific Computing (SciComp)
- Life Science Facility (LSF)
- BioImaging Facility (BIF)
- Electron Microscopy Facility (EMF)
- Austrian Science Fund (FWF) [P33367] Funding Source: Austrian Science Fund (FWF)
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.
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