3.1. Tissue Preparation
1. Fix the tissues in 10% (v/v) buffered formalin (see Chapter 47) for at least 24 h (see Notes 2 and 3).
2. Trim the tissues to the required size and to the region of interest (5).
3. Place the tissues in the automatic tissue processor. Tissues can be manually taken through the same series of solutions, with constant agitation throughout. The following regime should be employed: Reagent Routine (total 18 h, overnight)
50% (v/v) alcohol 2 h 70% (v/v) alcohol 2 h 95% (v/v) alcohol 2 h 100% (v/v) alcohol 2h 100% (v/v) alcohol 2 h Toluene 2 h
Toluene 2 h
Paraplast 2 h
In most machines, the beakers are static and hold the processing fluids, whereas the tissues are held in individual containers or cassettes in a suspended basket, and mechanically moved from one beaker to another. Agitation is provided by vertical or rotating movements of the basket. Tissues can be routinely processed overnight so that they are ready for embedding the next morning.
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