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
For clinical purposes it is often necessary to examine urine, fæces or
vomited matter for bacilli. Films are prepared in the usual way and
allowed to evaporate slowly, and then fixed by passing through the
flame, and then washed in distilled water before staining. In the case
of vomited matter and fæces this is usually done without difficulty.
In the case of urine however it is often difficult to get the urine
to evaporate completely. A syrupy layer remains, and if more heat be
applied it decomposes and chars, and the products cause precipitation
of aniline during subsequent staining processes. This may be partly
avoided by gently washing the film in distilled water before staining.
Another plan is to mix the urinary deposit with a little gelatine free
from organisms, such as that in unused culture tubes. The gelatin is
liquefied by heat, and mixed with the deposit. Films are made from this
mixture, and allowed to set, and then thoroughly washed in distilled
water. The film is then dried thoroughly, and the cover-glass laid flat
with the film uppermost, and a few drops of the staining fluid filtered
on to it. After it has been stained sufficiently the stain is drained
off, and the slip gently washed. The film may then be stained with some
contrast stain in exactly the same way as sections, again washed, dried
between folds of blotting paper, and mounted in balsam.
It is sometimes difficult to tell which is the side of the cover-glass
which bears the film. This is readily done by holding the glass
obliquely so that light from a window is reflected from its surface.
The side which is coated appears dull; while the other is smooth and
bright.
METHODS OF EXAMINING BLOOD.
In all these methods blood is obtained by pricking the skin of one of
the fingers, or the lobule of the ear, preferably the latter. The
skin must previously be washed with soap and water or ether, to remove
any grease or epithelial scales. The puncture should be made firmly so
that blood may escape freely. The finger or ear must not be squeezed.
Specimens must be made rapidly before red corpuscles have run into
rouleaux. The slides and coverslips employed must be scrupulously
clean, or it is impossible to get really good films. They should be
cleaned with nitric acid and alcohol according to the directions on
page 57.
Fresh specimens should be examined. The coverslip is made just to touch
the drop of blood at one edge, so as to transfer a small quantity
only, and is at once lowered on to the slide with the aid of a mounted
needle. If slide and coverslip be perfectly clean the blood will spread
out into a thin film, the corpuscles lying quite flat. If there be any
delay, or if the cover-glass be not quite clean the red corpuscles
will run into masses and the specimen will be useless for minute
examination. Another specimen may be mixed with a little of Ferrier’s
solution (p. 129) before mounting. Permanent coverslip films may also
be prepared.
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