The Elements of Bacteriological Technique: A Laboratory Guide for Medical, Dental, and Technical Students. Second Edition Rewritten and Enlarged.Eyre, J. W. H. (John William Henry)
Science
The Elements of Bacteriological Technique: A Laboratory Guide for Medical, Dental, and Technical Students. Second Edition Rewritten and Enlarged.
Eyre, J. W. H. (John William Henry)
Bacteriology -- Technique
4. Impregnate the tissues with mucilage for twelve to twenty-four hours,
according to size. Transfer the pieces of tissue to a bottle containing
sterilised gum mixture.
~Formula.~--
Gum arabic 5 grammes
Saccharose 1 gramme
Boric acid 1 gramme
Water 100 c.c.
5. Place the tissue on the plate of a freezing microtome (Cathcart's is
perhaps the best form), cover and surround with fresh gum mixture;
freeze with ether, or for preference, carbon dioxide, and cut sections.
6. Float the sections off the knife into a glass dish containing tepid
water and allow them to remain therein for about an hour to dissolve out
the gum.
(If not required at once, store in 90 per cent. alcohol.)
7. Transfer to a glass capsule containing the selected staining fluid,
by means of a section lifter.
8. Transfer the sections in turn to a capsule containing absolute
alcohol (to dehydrate) and to one containing xylol or oil of cloves (to
clear).
9. Mount in xylol balsam.
_Alternative Rapid Method._--
1. Cut very small blocks of the tissue.
2. Fix in formalin 10 per cent. aqueous solution (fixation
fluid No. 7, page 82) for 24 hours.
3. Transfer block to plate of freezing microtome and freeze
with carbon dioxide vapour.
4. Float the sections off the knife into a glass dish of
tepid water.
5. Stain the sections in glass capsules containing selected
stains.
6. Place the stained section in a dish of clean water and
introduce a glass slide obliquely beneath the section; with
a mounted needle draw the section on to the slide and hold
it there; gently remove the slide from the water, taking
care that any folds in the section are floated out before
the slide is finally removed from the water.
7. Drain away as much water as possible from the section.
Drop absolute alcohol on to the section from a drop bottle,
to dehydrate it.
8. Double a piece of blotting paper and gently press it on
the section to dry it.
9. Drop on xylol to clear the section.
10. Place a large drop of xylol balsam on the section and
carefully lower a cover-glass on to the balsam.
PARAFFIN METHOD.
1. ~Fixation.~ Place the pieces of tissue, resting on cotton-wool, in a
wide-mouthed glass bottle. Pour on a sufficient quantity of the
corrosive sublimate fixing fluid; allow the tissue to remain therein for
twelve to twenty-four hours according to size.
2. Pour off the fixing fluid and wash thoroughly in running water for
twenty minutes to half an hour to remove the excess of corrosive
sublimate.
[Illustration: FIG. 72.--~L~-shaped brass moulds.]
[Illustration: FIG. 73.--Paraffin kettle.]
3. ~Hardening.~ Place the tissues in each of the following strengths of
alcohol in turn for from twelve to twenty-four hours: 50 per cent., 75
per cent., 90 per cent., absolute.
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