Chapter 14 Fixation of Tetrahymena Cells for Electron Microscopy

Abstract

Fixation and preparation of Tetrahymena cells for electron microscopy is similar to procedures used for most cells. As with all fixation procedures, one should use the method most suited for fixation of the structures in which one is interested. This chapter describes the methods that are the best for routine preservation of the cilia, cortex, and cytoplasm. They can be carried out in any conventional laboratory equipped for electron microscopy. The same procedures are used to fix and stain cells for scanning electron microscopy (SEM) and for transmission electron microscopy (TEM). For SEM, unfixed cilia, pellicles, or other organelles are attached to polyethylenimine-coated coverslips and the coverslips are processed through the fixation and drying procedures followed by critical point drying. For TEM, organelles are pelleted and the pellets are fixed using the conventional fixation and embedding procedures, with the exception that pellets are generally embedded in flat molds.