The Microscope. Its History, Construction, and Application 15th ed.: Being a familiar introduction to the use of the instrument, and the study of microscopical scienceHogg, Jabez
History
The Microscope. Its History, Construction, and Application 15th ed.: Being a familiar introduction to the use of the instrument, and the study of microscopical science
Hogg, Jabez
Microscopy; Natural history
_Eosin stain_ is the most useful for sieve-tubes and plates. Make a
strong solution of eosin in equal parts of water and alcohol, and stain
the section for five or ten minutes. Wash well in methylated spirit,
dehydrate, clean in oil of cloves, and mount in Canada balsam.
_Bleaching Process._--The bleaching and clearing of vegetable
structures before staining is a very necessary process, especially
so if starch be present in any quantity. Clearing agents are of
two kinds--those which act by virtue of their property of strongly
refracting light, and those which disintegrate and dissolve the amyloid
cell contents. To the first class belong the essential oils, as oil
of cloves, Canada balsam, glycerine, and other similar bodies; to the
second class, solutions of potash, phenol, and chloral hydrate. The
actual value of some of these agents is questionable. The process
usually preferred is as follows: Place the sections in a fresh clear
solution of chlorinated lime, allowing them to remain until quite
bleached, say from two to four or five minutes; then gently warm in
a test-tube for a few seconds, and quickly replace the solution with
distilled water and boil for two or three minutes; repeat the treatment
with boiling water three times; wash with a one per cent. solution of
acetic acid, and finally with distilled water. The sections are then
quite ready for staining operations.
When the stem is hard and brown, a solution of chloride of lime should
be used--a quarter of an ounce of chloride dissolved in a pint of
water, well shaken and stood by to settle down, then pour off the clear
fluid for use. For hard tissues this solution answers well, but it is
not suitable for leaves, as they require not only bleaching, but the
cell contents should be dissolved out to render them transparent. A
solution of chlorinated soda answers well for both stems and leaves. It
is prepared as follows:--
To one pint of water add two ounces of fresh chloride of lime, shake
or stir it well two or three times, then allow it to stand till the
lime has settled. Prepare meanwhile a saturated solution of carbonate
of soda--common washing soda. Now pour off the clear supernatant
fluid from the chloride of lime, and add to it, by degrees, the soda
solution, when a precipitate of carbonate of lime will be thrown down;
continue to add the soda solution till no further precipitate is
formed. Filter the solution, and keep it in a well-stoppered bottle in
the dark, otherwise it speedily spoils.
Sections bleached in chlorinated soda must, when white enough, be
washed in distilled water, and allowed to remain in it for twenty-four
hours, changing the water four or five times, and adding a few drops
of nitric acid, or at the rate of eight or ten drops to the half-pint,
to the water employed before the final washing takes place. From water
transfer them to alcohol, in which they must remain for an hour or more.
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