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Methodology Guidelines
Special Considerations
Cell Adhesion
Cytology
100
Manual de operaciones de la Cytospin 4
solutions for extended times. These specimens may be so
rigid that it is difficult to get them to flatten on the slide.
The addition of a small amount of glycerol to the specimen,
allowing some 'soak' time, followed by use of the Cytospin,
will usually result in a reasonable preparation.
Many laboratories prefer to fix all specimens during
preparation. The usual protocol is to concentrate the
specimen, then re-suspend in an equal volume of Cytospin
Collection Fluid. For samples that must be diluted, the
diluent can be fixative. These fixation steps are generally
done just before adding the sample to the Cytospin sample
chamber assemblies.
Whether the specimen has been fixed before deposition on
slides or not, immediately after removing the slides from
the Cytospin, they should be immersed in 95% alcohol to
complete fixation and dehydration. Since the cells will still
be wet, and will not have become totally bound to the glass
slide, use care in this transfer. (Ease the slide into the fixative
in the container). Many complaints of poor cell capture can
be traced to lack of care in this step of the procedure.
Successful application of the Cytospin requires that cells
adhere to the glass slides. For many routine applications, it
is sufficient to use clean slides. Slides may be cleaned using
alcohol. The increasing use of long staining techniques, such
as immunostaining, may require additional ways to ensure
adhesion of samples. The use of coated slides will increase
cell retention and reduce the incidence of 'floaters' in the
subsequent staining baths.
The majority of the procedures previously discussed apply
to cytological specimens. However, there are a number
of specific specimen types that require special processing.
Often, the cytology specimen will contain clots, fibrin
webs, or tissue fragments. These will all interfere with the
Cytospin preparations. Small floating clots and fragments
should be removed with forceps. These may be saved for
cell block procedures. Fibrin webs are generally too friable
A78310250ES 5.1ª Edición
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