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CryoCycle your grids: Plunge vitrifying and reusing clipped grids to advance cryoEM democratization.

ABSTRACT: CryoEM democratization is hampered by access to costly plunge-freezing supplies. We introduce methods, called CryoCycle, for reliably blotting, vitrifying, and reusing clipped cryoEM grids. We demonstrate that vitreous ice may be produced by plunging clipped grids with purified proteins into liquid ethane and that clipped grids may be reused several times for different protein samples. Furthermore, we demonstrate the vitrification of thin areas of cells prepared on gold-coated, pre-clipped grids.

SUBMITTER: Serbynovskyi V

PROVIDER: S-EPMC11247934 | biostudies-literature | 2024 Jul

REPOSITORIES: biostudies-literature

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CryoCycle your grids: Plunge vitrifying and reusing clipped grids to advance cryoEM democratization.

Serbynovskyi Viacheslav V Wang Jing J Chua Eugene Yd EY Ishemgulova Aygul A Alink Lambertus M LM Budell William C WC Johnston Jake D JD Dubbeldam Charlie C Gonzalez Fabio A FA Rozovsky Sharon S Eng Edward T ET de Marco Alex A Noble Alex J AJ

Research square 20240702

CryoEM democratization is hampered by access to costly plunge-freezing supplies. We introduce methods, called CryoCycle, for reliably blotting, vitrifying, and reusing clipped cryoEM grids. We demonstrate that vitreous ice may be produced by plunging clipped grids with purified proteins into liquid ethane and that clipped grids may be reused several times for different protein samples. Furthermore, we demonstrate the vitrification of thin areas of cells prepared on gold-coated, pre-clipped grids ...[more]

PMID: 39011111

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