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A primary mission of the Advanced Electron Microscopy Facility is to develop techniques for preserving cellular structure with the highest degree of reliability. These techniques involve different methods for rapidly freezing our samples in order to halt structural and biochemical activity in a very short timeframe, thus preserving structure in the "live" state. Once the sample is preserved in the "live" state, it is then possible to study the ultrastructure of these samples using not only basic Electron Microscopy imaging techniques, but also state-of the-art techniques such as: 1) 3-D electron tomography, and 2) immuno-cytochemistry.
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03/02/09 06:51am
3-D Electron Tomography
09/24/08 03:12pm
Why use Cryo-Preservation Techniques to Study the Ultrastructure of Biological Material
Rapid freezing and freeze-substitution are preferable to standard, room-temperature chemical fixation techniques for several reasons. First, rapid-freezing brings biological activity uniformly to a halt within a matter of milliseconds. Chemical fixation, on the other hand, is progressive and takes many minutes or hours to halt cellular processes. Since many cellular events are very rapid (i.e. vesicle budding in the Golgi apparatus) the chemicals cannot stop everything all at once, thus the cell cannot be studied in its truly native state. Second, freeze-substitution allows for the removal of water (dehydration) and infusion of fixatives while the cells is still in a frozen state. This means that the artifact introduced by multiple changes of increasing concentrations of solvents at room temperature is avoided.
09/24/08 03:11pm