"Tips and tricks" for high-pressure freezing of model systems.
High-pressure freezing (HPF) has been around since the mid-1980s as a cryopreparation technique for biological electron microscopy. It has taken quite some time to "catch on" but with the recent interest in cellular tomography and electron microscopy of vitreous cryosections it has been used more frequently. While HPF is relatively easy to do, there are a number of steps, such as loading the sample into the specimen carrier correctly, that are critical to the success of this method. In this chapter we discuss some of the "little" things that can make the difference between successful or unsuccessful freezing. We cover all aspects of HPF, from specimen loading to removing your sample from the carriers in polymerized resin. Our goal is to make it easier and more reliable for HPF users to get well-frozen samples for their research.
McDonald
Kent
K
Electron Microscope Laboratory, University of California, Berkeley, California 94720, USA.
Schwarz
Heinz
H
Müller-Reichert
Thomas
T
Webb
Rick
R
Buser
Christopher
C
Morphew
Mary
M
D000818:Animals; D015925:Cryopreservation / Q000295:instrumentation / Q000379:methods*; D008854:Microscopy, Electron / Q000295:instrumentation / Q000379:methods*; D008954:Models, Biological; D011312:Pressure; D013194:Staining and Labeling / Q000379:methods; D016707:Tissue Fixation / Q000295:instrumentation / Q000379:methods