Cryotechniques

Location: Thomas Building, DE-780
Contact phone: (206) 667-4289
Contact fax: (206) 667-1166
Contact e-mail: emsr@fredhutch.org

Electron microscopy owns several pieces of Leica cryo equipment including a High Pressure Freezing apparatus, freeze substitution system, and low temperature cryochamber. This equipment produces superior cellular preservation for morphological studies and better antigen retention for immunolabeling work.

High Pressure Freezing (HPF)

The Leica EM PACT2 is a high pressure freezing machine that is designed to freeze samples up to 200-300 um into the specimen without significant ice crystal damage. HPF provides for better antigen retention for immunolabeling. HPF is followed by freeze substitution or cryo-ultramicrotomy.

Automatic Freeze substitution (AFS)

Automatic Freee Substitution (AFS) chamberThe Leica EM AFS2 is capable of freeze substitution, progressive lowering of temperature techniques, and low temperature embedding and polymerization of resins for improved preservation of ultra structure and antigenicity.

Cryo-microtomy

Cryo-ultramicrotomy is performed by the Leica EM FC6 cryochamber. It is designed for low temperature sectioning of samples at temperatures from -15 to -185 degrees C.

More information on the equipment is found at the Leica web site.

Leica VT1200 S Fully automated vibrating blade microtome:

VibratomeThe following instruments are available in the Electon Microscopy core facility for use in sample preparation for electron microscopy.

The Leica VT1200 S fully automated Vibratome uses a vibrating razor blade to cut sections from 1-999 microns under physiological conditions without freezing or embedding, and thus ultrastructure is well preserved.  The tissue can be fixed or fresh when using a vibratome.  The vibratome maintains cell morphology, enzyme activity and cell activity.  The microtome is well used for immunocytochemistry and histoenzymology. 

Ultimately, vibratome tissue sections can be trimmed down and used for high pressure freezing.

Located in the South Lake Union neighborhood of Seattle, the facility serves Fred Hutchinson, Cancer Consortium, and all other Seattle area researchers.