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
Cellulose, as it occurs in plant life, presents a variety of physical
properties: sometimes it is soft, as in young plants, and again quite
dense in older structures. This fact accounts for the varying results
obtained when cellulose is subjected to the action of staining fluids,
and whether the cellulose occurs in a nearly pure form, as in cotton
fibre, or in the modified form of lignine or woody-fibre. Stains
which readily attack young tissue have little or no effect upon it in
its maturer form. It is of much importance, then, in the staining of
fibres, as well as sections for the microscope, that the cellulose
should take the stain uniformly.
The staining of tissues may be effected in four ways. First, when the
stain has sufficient affinity for the tissue to be retained by it
without the intervention of any outside agent. Second, when the stain
and mordant are mixed and applied to the tissue in one solution. These
two are the simplest and easiest methods of staining. Third, when the
tissue is first immersed in the staining liquid and then transferred to
some other liquid which shall fix the colour upon the tissue. Fourth,
when the tissue is first impregnated with the mordant, or fixing agent,
and then immersed in the stain. The last method is the one usually
followed in commercial works, and it is to be recommended in the
staining of microscopical preparations which do not readily take the
stain.
_Nuclear Stains._--As in both vegetable and animal sections it is
generally the nuclei which form the landmarks of the structure, so the
most important class of reagents which are used in any of the branches
of microscopical work are the “nuclear stains.” There are several of
these stains, the most important of which is the hæmatoxylin, and
when proper solutions are used the results are very satisfactory.
Many formulæ have been given, but there are three only reliable,
Delafield’s, Kleinenberg’s, and Ehrlich’s, in all of which alum is
present as an ingredient; the idea being that the alumina forms with
the colouring matter an insoluble lake, and so acts as a mordant.
In _Delafield’s_ solutions a large proportion of alum to hæmatoxylin
is used, and methylic alcohol (wood-spirit in the place of rectified
spirit).
For _Kleinenberg’s_ solution many different formulæ exist. Squire’s
improved formulæ for both stains is given in the Appendix, “_Formulæ
and Methods_.”
Hæmatoxylin solutions stain the nuclei violet, and in order to change
this into blue, the sections should be transferred to water taken from
the house supply, not distilled water; but as the alkalinity of the
water varies in different localities, a better and more uniform result
is obtained by using a weak solution of bicarbonate of sodium (half a
grain to the ounce).
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