Sample preparation

Sample preparation

Background

Samples for cryo-EM are prepared by vitrification: cryo-fixation by rapid freezing. These frozen samples are then imaged in a near-native state at liquid nitrogen temperature. Because the samples are not chemically fixed or stained they can be imaged the highest resolution, but are also very sensitive to electron beam damage. Therefore pictures are taken at a very low electron dose and with the best imaging cameras, in order to prevent radiation damage and get a good signal. 

Instruments

NeCEN is equipped with a FEI Vitrobot for plunge freezing of biological samples. For challenging samples, we offer assistance during buffer optimization and sample screening. For this a basic equipped wet-lab and dedicated entry level screening TEM are available on site.

Sample preparation and quality assessment

To obtain grids of sufficiently quality for data collection, the NeCEN staff will freeze and load batches of 8 grids. A maximum of 2 times 8 grids per day can be screened.

Sample preparation before data collection

To freeze samples before data collection session, the NeCEN staff will freeze and load batches of 8 grids.