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

TL;DR

This paper introduces a method called CryoCycle to reduce costs in cryoEM by reusing clipped grids for vitrifying and imaging different protein samples.

Contribution

The novel contribution is a reliable method for blotting, vitrifying, and reusing clipped cryoEM grids for multiple protein samples.

Findings

Vitreous ice can be produced by plunging clipped grids with purified proteins into liquid ethane.

Clipped grids can be reused several times for different protein samples.

Thin areas of cells can be vitrified on gold-coated, pre-clipped grids.

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.

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