Preparation of Freeze-Dried Specimens for Electron Microscopy
The application of freeze-drying to electron microscopy (EM) involves rapid freezing of the specimen followed by the removal of the frozen water by controlled sublimation. In the simplest case, freeze-drying is followed directly by observation in a transmission electron microscope. The freeze-drying step can also be incorporated into a wide range of more complex processing protocols, which we will refer to briefly, directing the reader to recent reviews and some examples from the literature. Our emphasis will be on the methodology of freeze-drying itself and on the results that can be expected from this technique. Today there are many preparative techniques to choose from, including critical point-drying, freeze-substitution, freeze-etching, and standard fixation and embedding. All are capable of yielding excellent morphological preservation when appropriately applied. We will discuss freeze-drying in the context of these methods in order to identify its unique capabilities.
KeywordsRapid Freezing Freezing Damage Freezing Technique Electron Microscope Autoradiography Electron Microscopy Grid
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