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Approaches to Using the Chameleon: Robust, Automated, Fast-Plunge cryoEM Specimen Preparation.

Authors :
Levitz TS
Weckener M
Fong I
Naismith JH
Drennan CL
Brignole EJ
Clare DK
Darrow MC
Source :
Frontiers in molecular biosciences [Front Mol Biosci] 2022 Jun 23; Vol. 9, pp. 903148. Date of Electronic Publication: 2022 Jun 23 (Print Publication: 2022).
Publication Year :
2022

Abstract

The specimen preparation process is a key determinant in the success of any cryo electron microscopy (cryoEM) structural study and until recently had remained largely unchanged from the initial designs of Jacques Dubochet and others in the 1980s. The process has transformed structural biology, but it is largely manual and can require extensive optimisation for each protein sample. The chameleon instrument with its self-wicking grids and fast-plunge freezing represents a shift towards a robust, automated, and highly controllable future for specimen preparation. However, these new technologies require new workflows and an understanding of their limitations and strengths. As early adopters of the chameleon technology, we report on our experiences and lessons learned through case studies. We use these to make recommendations for the benefit of future users of the chameleon system and the field of cryoEM specimen preparation generally.<br />Competing Interests: Authors MD and IF were employed by the company SPT Labtech. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.<br /> (Copyright © 2022 Levitz, Weckener, Fong, Naismith, Drennan, Brignole, Clare and Darrow.)

Details

Language :
English
ISSN :
2296-889X
Volume :
9
Database :
MEDLINE
Journal :
Frontiers in molecular biosciences
Publication Type :
Academic Journal
Accession number :
35813832
Full Text :
https://doi.org/10.3389/fmolb.2022.903148