WEHI: Construction and testing of a plunge freezing device for cryo-electron microscopy (2021-22)
Construction and testing of a plunge freezing device for cryo- electron microscopy at WEHI (2021-22)
Proteins and viruses intended to be visualised by cryo- electron microscopy are prepared by rapid freezing in liquid ethane. Using supplied components and plans, the team will assemble and program a vitrification device for routine use by WEHI structural biologists. The design comprises a published linear solenoid and piezo nebuliser design, and will be complemented by a temperature-controlled cryostat, with plans for the latter supplied by the Laboratory of Molecular Biology at the University of Cambridge. The project therefore involves two parts that can be considered separately. Part 1 is construction of the cryostat and control interface to maintain liquid ethane at a temperature close to its melting point (https://aip.scitation.org/doi/full/10.1063/1.4967864). Part 2 is construction of a combined spray freezer and mechanism to accelerate the sample into liquid ethane. Students are welcome to study the plans for the prototype (https://journals.iucr.org/d/issues/2019/12/00/ih5001/index.html). The Supplementary Material in this article provides a link to the Github site with the control scripts. A budget for the requisite materials has been approved. All materials and spares will be available well in advance of project start.
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