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
In making up these solutions care must be taken that _only distilled
water_ is used, and that all the vessels employed have been previously
rinsed out with it, otherwise precipitation of the hæmatoxyline will
occur.
Should sections be overstained in hæmatoxyline, this may be remedied
by washing it in a half per cent. solution of acetic acid, until
sufficient of the stain is discharged, but the staining is more diffuse
than if the happy mean had been hit in the first instance.
Hæmatoxyline stains the nuclei of the cells a beautiful violet colour,
and also tints, more or less lightly, the cell protoplasm and the
fibrous elements. It also stains the axis cylinders of nerves, and is
much used in special staining of the nerve centres as will be described
later, (pp. 88–91).
The stain is permanent. Sections may be mounted either in Farrant’s
solution, or in Canada balsam, the latter being preferable.
*Eosine.*--Much more satisfactory results are obtained from the
commercial eosine (an amorphous orange powder used in dyeing and in the
manufacture of red ink), than from the pure crystalline form.
It may be used as an aqueous solution (1/30 per cent.) or as a
solution in absolute alcohol (1/15 per cent.). Sections stained in the
former should be rapidly passed through a one per cent. solution of
acetic acid in order to “fix” the stain, and then washed in distilled
water.
It is a very transparent stain, and the most delicate details of a
section stained with it are perfectly visible.
It stains the nucleus but slightly, while it stains the cell protoplasm
and fibrous tissues and especially muscular tissues a beautiful rose
colour.
It will be seen, therefore, that it stains those parts which are left
unstained by hæmatoxyline, and _vice versâ_. This complementary action
is applied in the following method.
*Double staining with eosine and hæmatoxyline.*--Sections having been
stained in hæmatoxyline in the ordinary way, are washed in distilled
water, and dehydrated in a solution (about 1 in 1500) of eosine in
absolute alcohol. They should remain in this for about two minutes, and
then be passed through oil of cloves and mounted in Canada balsam in
the ordinary way.
This method gives extremely useful and beautiful results with almost
all tissues, and is superior to picrocarmine for differentiating
the tissue elements. Thus, the nuclei are stained violet, the cell
protoplasm a much paler and warmer violet, the fibrous tissues pink,
and red blood corpuscles orange or brick red.
The alcoholic solution of eosine is also used as a contrast stain after
staining for micro-organisms with blue or violet dyes.
*Carmine.*--It is made as follows:--
Carmine (best) 2 1 drachm.
Strong ammonia 2 1 drachm.
Distilled water 100 6 ounces.
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