Section Cutting and Staining: A practical introduction to histological methods for students and practitionersColman, Walter S.
Science
Section Cutting and Staining: A practical introduction to histological methods for students and practitioners
Colman, Walter S.
Microscopy
The process of *clarifying* must be performed in some medium in which
Canada balsam is readily soluble, and which is also readily miscible
with alcohol. Those most frequently employed are oil of cloves, xylol,
oil of bergamot, oil of cedar, and origanum oil. The first named has
always been much used because of its agreeable odour, its cheapness,
and the ease with which it can be obtained. But it has the disadvantage
of dissolving out many important staining reagents, especially eosine
and the various aniline colours. In addition as it dissolves celloidin,
sections cut in this medium tend to fall to pieces when transferred to
oil of cloves, and one of the other oils (which have no solvent action
on celloidin) should always be employed with celloidin sections. Oil
of bergamot is the most generally useful, but rather expensive. Where
there are special reasons for employing other dehydrating agents, they
will be indicated in the special directions for particular staining
methods in Chapters VI. and VII.
As soon as the section is plunged into the oil, the alcohol rapidly
diffuses out, so that the edges of the section fly out with it, and
the section floats quite flat on the surface of the oil. When it is
completely clarified (in about a minute), as shown by its sinking in
the oil, it should be transferred to the slide by the section lifter,
and the oil drained off. Excess of oil may be removed by pressing
gently on the section with a flat piece of filter paper folded several
times. If carefully performed this manœuvre will not injure the
section, but it requires practice.
If the tissue is very delicate, and likely to be injured by changing
from one vessel to another, or if it is larger than the section lifter
will conveniently carry, it should be floated out on a glass slide,
and, as much water as possible having been removed by blotting paper,
should be dehydrated by adding a little alcohol from a pipette once or
twice. Most of the alcohol should then be removed by tilting the slide,
and before the remainder has evaporated, some oil of cloves or bergamot
should be added from another pipette. The section will float on the oil
at first, but the latter will gradually come through and appear on the
top of the section. When this occurs the clarification is complete, and
the oil may be run off by tilting the slide and the section mounted in
Canada balsam.
*Cementing of cover-glasses.*--The cover-glasses may be cemented down
to prevent their shifting and spoiling the specimen. If the cover-glass
be circular, a Shadbolt’s turntable should be used. It consists
simply of a horizontal heavy brass disc, rotating easily on a pivot.
There are a number of circles traced on the disc concentrically. The
slide is then fixed on the disc by means of the clips, so that the
circumference of the cover-glass corresponds to one of the circles.
The disc is then rotated and the cement applied to the edge of the
cover-glass with a brush.
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