Electron Cryomicroscopy of Fibrillar Collagens - Methods in Molecular Biology: Extracellular Matrix Protocols - page 99

Fig. 5. Schematic representation of the first-stage of sample preparation of the ice
wedge prior to freezing.

Fig. 6. Schematic representation of the Cressington CFE-50C instrument used here
for freeze-fracture, etching, and low-temperature shadowing. The system allows fine
temperature control (down to -204 ° C) of the specimen and well-controlled evapora-

tion of the shadowing metal.

4. Raise the microtome slightly and then position it over the top of the sample to

protect it from contamination during freeze-etching.

5. Set the thermal control of the stage heater to -100 ° C and turn on the heater. Once

the stage temperature reaches -100 C etch the sample for 3 min and then turn off

the heater, allowing the stage to return to -189

°

° C.