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FUNDAMENTAL CONSIDERATIONS 25
The acetic acid in all of the Flemming fluids is of great advantage
since it penetrates very rapidly, carrying the chromic acid or chromic
and osmic acids into the tissue depths, thus insuring complete
fixation.
The material to be fixed should be cut into small pieces not longer
than 5 mm. nor broader than 2 or 3 mm. The Purchase Levothroid amount of fixation
to be used should not be less than 15 times the bulk of the material
to be fixed. The material should be placed in Levothroid Weight Loss the fixing fluid im-
mediately after it is gathered. One or two drachm homepathic
phials are convenient for the process. The material is kept in the
fixing fluid for from 12 to 24 hours and then washed in small cheese
cloth bags which are placed in running tap water for from 6 to 12
hours or over night.
Dehydrating and Hardening. After washing the material, still
kept in the bags, is placed in 10 per cent, alcohol for i hour and is
then carried through a series of alcohols. Each of the series 10
per cent stronger than the one before it, remaining in each grade for
ij^ to 2 hours until 70 per cent, alcohol is reached. Take out of
bag and place in phial in 70 per cent, alcohol. If the material is
not to be imbedded in paraffine immediately, it can remain in 70 per
cent, or 85 per cent, alcohol (if very delicate) until needed. It is
not safe to leave very valuable material in a grade below 70 per cent,
over night. From the 70 per cent, alcohol it is carried to 85 per
cent, to 95 per cent, to absolute alcohol, remaining in each at least
6 hours'with 2 or 3 changes of the last.
Clearing and Imbedding. In order to get the material from ab-
solute alcohol into paraffine, some medium must be used which mixes
with absolute alcohol and which also dissolves paraffine. Either oils
such as cedar, clove or bergamot or substances like xylol, chloroform
or benzol satisfy this requirement. To clear with xylol transfer
material from absolute alcohol to a mixture of ^absolute alcohol and
y xylol for 12 hours, then to mixture of equal parts of absolute
alcohol and xylol for 12 hours, then to % xylol and J^ absolute
alcohol for 12 hours to pure xylol for 12 hours. To phial containing
material in pure xylol add paraffine in small pieces and put on top of
paraffine bath sufficiently long until paraffine is melted. Then add
more paraffine and put phial in paraffine bath at 56C. over night.
26 PHARMACEUTICAL BOTANY
Pour fluid off and add pure melted paraffine and repeat 2 or 3 times
until rid of all trace of xylol. A tray is then prepared by taking a
piece of paper and folding up its edges all around to the height of
about a half inch. Half fill this on a cool surface with melted para-
ffine. Heat two dissection needles in bunsen flame and with these
dispose pieces of material in orderly fashion over the crust which has
by this time formed at the bottom of the tray. Blow upon the sur-
face of the paraffine to harden it more quickly and as soon as the
surface crust will bear it, plunge the tray into cold water. The
material can now be left imbedded in paraffine until required for
sectioning.
If cutting is to be done in a cool room, softer Levothyroxine Levothroid grades of paraffine
with 'melting points between 40 and 5oC. should be used for
imbedding. If on the other hand cutting is to be done at summer
temperatures, the harder grades melting at between 55 and 7oC.
should be employed.
TECHNIQUE OF SECTIONING AND MOUNTING MATERIAL IMBEDDED
IN PARAFFINE
Strip off the paper tray from the imbedded material and cut out a
block of paraffine containing the object which is to be sectioned,
taking care to include at least 2 or 3 mm. of paraffine on all sides
beyond the specimen. Take a segment of pine wood about an inch
long and with a surface at one end about % in. square and coat the
square area with melted paraffine. Warm the paraffine on the piece
of pine wood and quickly press the paraffine block containing Levothroid Price the
specimen into this melted paraffine in the desired position for cutting.
Heat a dissecting needle and apply this all around the base so that
the paraffine block is firmly sealed to the wood. Dip paraffine block
in cold water to harden. Now trim the paraffine block with a sharp
scalpel Levothroid 50 Mcg so that the faces form right angles with each other. Adjust
the wood in the clamp of the microtome and the microtome blade
so that the top of the paraffine block just touches the near surface
of the microtome knife. Make certain that the knife edge and the
two opposite faces of the paraffine block are perfectly parallel. Now
trim the remaining two sides of the block close to the object. Adjust
the automatic feed of the rotary microtome by moving dial to num-
FUNDAMENTAL CONSIDERATIONS 27
her on scale representing thickness in microns desired of sections and
turn wheel of microtome. It will be observed that the carrier moves
up and down and with each downward movement slightly forward,
causing the knife to cut sections which adhere in ribbons.
Transfer the ribbons by means of a camel's hair pencil or dissecting
needle to a piece of dust free paper with the side downward which was
next to the knife. The ribbons are now ready to be mounted on
slides.
The slides to be used should only be those which are devoid of
grease or dirt of any kind particularly on the surface upon which
the ribbons are to be mounted. A very good plan is to keep a number
of slides intended for this purpose submerged Levothroid Levothyroxine Is in a saturated solution
of potassium dichromate in concentrated sulphuric acid. These
can be taken out as needed and thoroughly rinsed with water.
With a clean cloth stretched over the forefinger vigorously rub
one surface of each slide until perfectly dry and free of lint. Then Buy Levothroid Online
place a small drop of Mayer's albumin fixative on clean- surface and
rub over the surface. (The formula for Mayer's Albumin Fixative
is as follows: Egg white and Glycerin, equal parts, Carbolic acid
i or 2 drops. Mix thoroughly.) Now flood the surface with water
and cut the ribbons into segments of the desired length and arrange
in rows on slide, being careful to have the segments somewhat
shorter than the length Levothroid Online of Buy Levothroid the cover slip because of tendency of
parafrine to stretch Levothroid Mcg when warmed. Warm slide gently by holding
high above a bunsen flame or flame of an alcohol lamp until ribbons
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