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Primary Techniques

The primary techniques used here are:

Using the facilities

FEI Vitrobot freezing sample in ethaneTrained researchers can use microscopes themselves. Alternatively, MEMC personnel can assist researchers in collecting images.

Please contact Michael Purdy (mpurdy@virginia.edu) to discuss how best to use the facility to aid your project. A project application will be required, at which time access to the scheduling system will be provided.

Clients will need to provide a 8-12Tb USB storage drive for multi-day data collection on the Titan Krios or download data from MEMC servers.  Storage of data will only be guaranteed for one month.  Images collected on the Spirit or F20 will accessible by download from a remote server.

Sample preparation

Researchers can freeze samples in liquid ethane with a manual guillotine or using an FEI Vitrobot. Several options are also available for negative-staining.

Typical sample requirements pre grid are 3 µL at concentrations of 0.1-1.0 mg/ml.  A minimum of 20 µL is preferred in order to freeze multiple grids.

Frozen Sample

By examining frozen, unstained samples in these high resolution transmission electron microscopes and taking images from different angles, researchers get structural data down to the molecular level, without having to crystallize proteins. This data complements the expertise in protein X-ray crystallography and NMR in the Department of Molecular Physiology and Biophysics