Cryo EM Sample Preparation

The imaging capabilities of the facility are supported by a complete pipeline of state-of-the-art sample preparation equipment for cryo applications.

Cryo samples are vitrified by automated plunge freezing (FEI Vitrobot) or by high pressure freezing (Baltec HPM010 or freezing after light-stimulation, Leica HPM100). Cryo-ultramicrotomy (also of vitreous samples) is done in an area of controlled temperature and humidity at a Leica FC6 ultramicrotome. On the light microscopy site, sample preparation and correlative work is supported by a Zeiss LSM 700 confocal microscope which has a cryostage attached to enable visualization of vitrified samples by fluorescent light.

Various general EM sample preparation equipment is available. Room Temperature Microtomy is done at a Leica UC7 ultramicrotome or a Reichert Ultracut S.

Identification of fluorescent structures down to 1.1 micron immediately before high pressure freezing is done at a fully apochromatic corrected stereo microscope with 16.5:1 zoom optics (Leica M165 FC stereo microscope). Carbon films of high quality and reproducible thickness are generated by a Cressington Carbon Coater type 208.

 

Poster Hiwis

Partners & Funding sources