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Methodology Guidelines
Samples which contain 'average' cells, that is cells with an
approximate diameter of ten to twelve microns, produce
excellent Cytospin preparations at cell densities of one
million cells (1x10
) per ml. Specimens containing large
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cells require lower cell concentrations, and specimens with
tiny cells, cell organelles, or bacteria, may require higher
concentrations. The absolute concentration required will
be somewhat dependent on the processing methodology
employed. As a general rule, the concentration chosen
should be such that the cells within the sample have
adequate space to spread into a monolayer on the slide
surface, with minimal overlap, or piling up of cells.
Ideally, the concentration should be high enough that
there is not too much space between cells. Having
sufficient concentration of cells speeds up evaluation of the
preparation, since little time will be lost in searching for cells
to evaluate.
A quick method for approximating the number of cells
present in a sample is to place a single drop of the sample
on a slide and cover with a 24 x 50 mm coverslip. By
lowering the condenser of the microscope, or by closing the
microscope condenser diaphragm, the unstained cells can
be seen (although detail will not be seen). Using the 10 x
objective, scan the field and pick an area that appears about
average for the entire slide. The cells will mostly likely not
be evenly spread, which is why it is necessary to select an
average area. Now switch to the 40 x objective. You may also
need to open the diaphragm or raise the condenser slightly.
A78310250ES 5.1ª Edición
Manual de operaciones de la Cytospin 4
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