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
_Paraffin._--Paraffin is a very convenient embedding medium for
delicate structures, as very thin sections can be obtained and the
paraffin need not be removed from the section until the latter is
safely on the slide. It is unsuitable for large sections. Staining
operations are not easily carried out after cutting in paraffin, and
it is better to stain the blocks of tissue in bulk before embedding.
The best stains for penetrating are borax carmine (p. 75), alum carmine
(p. 76), and Kleinenberg’s hæmatoxyline (p. 70). The tissue must be
left in them for four to ten days.
Various kinds of paraffin are employed. It is usual to keep two kinds,
one “soft,” melting at 110° F., and another “hard,” melting at 140° F.
A mixture of two parts of the hard and one of the soft will be found
most generally useful. In winter a large proportion of the soft variety
and in hot weather a larger proportion of the hard may be required. A
paraffin mass which is always available has been suggested recently by
Dr. F. E. Batten, who employs an ordinary white candle, composed of
paraffin and wax. If the mass is found to be too hard, it can easily be
made of a suitable consistence by adding a little paraffin with a low
melting point.
To prepare a piece of tissue for embedding in paraffin, it should be
stained, washed in distilled water, and as much moisture as possible
removed by blotting paper. The block is then dehydrated, first in
methylated spirit for several hours, finally in absolute alcohol. It
is taken carefully by means of forceps from the alcohol and placed
in xylol for an hour or two according to size. Superfluous xylol is
removed from the surface, and the tissue placed in the melted paraffin.
This will set round the cold tissue at once, but soon melts again and
must be kept at a temperature just above melting point for one to
four hours, according to size. The tissue is then transferred to a
mould (which can be easily made of paper), about half an inch cube,
and melted paraffin poured round it until the mould is full. The mould
may be made by folding a piece of paper to form a box about half an
inch cube, or a small pill box may be used. Another convenient method
is to place two L-shaped pieces of lead in contact with each other so
as to enclose a space of suitable size as in the diagram (fig. 3). The
tissue is now hermetically sealed, and can be kept indefinitely if it
is not convenient to cut it at the time. To prepare it for cutting,
all superfluous paraffin is trimmed away with a warm knife, and the
block is fixed on a piece of wood, cut so as to suit the clamp of the
microtome, by melting the lower end of the paraffin block with a hot
needle or wire and pressing it down on the wood.
[Illustration: Fig. 3.]
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