Paraffin is the agent usually employed for embedding purposes. Melt it, and add a little lard to soften it; the addition of a little clove oil renders it less adhesive.
Melt the paraffin at as low a temperature as possible, and pour it into a paper cone. Dip the object into this and remove immediately; as soon as the layer of paraffin surrounding it becomes hardened, replace it in the paraffin; this prevents overheating the tissues.
Where the tissues are too soft to be cut, they may be soaked in a solution of gum arabic and dried; in this condition they can be readily cut, after which the gum can be dissolved off. This is an extremely useful method for cutting the lung or other organs where an interstitial support is needed. For a very thin object, a cork fitting any kind of a tube is to be split, and the object placed between the two parts; the cork is then thrust into the tube, and a sufficient degree of firmness will be obtained to allow cutting. The sections should always be manipulated with camel's hair brushes.
Much practice will be required before dexterity is attained.
All water must be removed from the tissue, either by drying or by immersing it in rectified spirits, and then in absolute alcohol, and the alcohol driven off by floating it upon oil of clove or turpentine. The substances used to preserve the tissues are Canada balsam, Dammar balsam, glycerine, Farrant's solution, potassium acetate, spirits, naphtha, and creosote.
The section is to be floated on to the slide or placed in position with a camel's hair brush. It should be spread out, and then examined under the microscope for the purpose of improving its position if necessary, or of removing any foreign particles. A drop of the preserving medium is then placed upon it, and another placed on the cover and allowed to spread out. The cover is then taken by a pair of pincers and inverted over the object, and one edge brought to touch the slide at one part of its margin. The cover is then gently lowered, and the whole space beneath the cover filled and the tissue completely saturated. If air bubbles show themselves, raise the cover at one corner and deposit a further quantity of the medium.
The slide should be set aside for a few days. First, the excess of the medium must be removed; if it is glycerine, much of it can be removed by a piece of blotting paper, but the cover must not be touched, for it is easily displaced; that near the cover can be replaced by a camel's hair brush. A narrow ring of glycerine jelly should be placed around the edge of the cover, to fix it before the cement is applied. When this has set, a narrow strip of cement is to be put on, just slightly overlapping the edge of the cover and outside the margin of the jelly. Until it has been perfectly secured, a slide carrying glycerine must never be placed in an inclined position, as its cover will slide off.
Canada balsam may be prepared as follows: Place some pure Canada balsam in a saucer, and cover with paper to exclude dust; dry it in an oven at a temperature of 150°; when it cools, it will become hard and crystalline. Dissolve this in benzole, and use in the same way as glycerine.
Dammar is now used as a substitute for Canada balsam. By its use the tissues are rendered more transparent. To prepare it, dissolve one-half ounce of Dammar rosin and one-half ounce of gum mastic in three ounces of benzole, and filter. This may be used to mount unsoftened bone and tooth, hair, brain, and spinal column, and most tissues that have been hardened in alcohol or chromic acid, which require to have their transparency increased.
Glycerine is not adapted for white fibrous tissue or blood vessels, unless they have been hardened in chromic acid, as it causes the white fibers to swell up and lose their normal features. Sections of liver, lung, skin, and alimentary canal show better in glycerine unless they have been stained.
Farrant's solution may be substituted for glycerine in many instances, because of its feebler tendency to render the tissues transparent. It consists of equal parts of gum arabic, glycerine, and a saturated solution of arsenious acid. In mounting preparations with this medium, the covered object should be allowed to lie a day before the varnish is applied, so that the cover may be fixed, and thereby prevented from being displaced. Rectified spirits may be used for mounting softened bone and tooth, and naphtha and creosote are useful for preserving urinary casts.
When the section is mounted in Canada or Dammar balsam, no cement is required, but for all other preservative media the margin of the cover must be covered with cement. To do this, dry the edges of the cover thoroughly with bibulous paper, and paint a layer of gold size, allowing it to overlap the cover an eighth or sixteenth of an inch, then cover this with white zinc cement.
To obtain a section of bone or tooth requires a grinding down of the tissue until it is so thin as to be transparent. A section should first be cut as thin as possible by a fine saw. It should be attached by the flattest side to a piece of glass, and then ground down by a grindstone or by very fine emery, on a perfectly flat piece of lead. When sufficiently thin and transparent, mount in rectified spirits or Dammar. Sections of the tongue may be made by embedding in paraffin, and mounted in Farrant's solution or glycerine.
Sections of the stomach may also be made by embedding in paraffin, but better ones can be made by freezing. Farrant's solution makes a good mounting.
The intestines also give a better section from freezing than by embedding, as the paraffin injures the villi; mount in the same medium as the stomach.
The liver may be embedded in paraffin, and the section mounted in Farrant's solution or glycerine. The kidney may be treated in the same way. The cornea of the eye can be readily cut by embedding in paraffin, and the section may be mounted in Farrant's solution. The crystalline lens and retina may be treated similarly.
The brain and spinal cord should be embedded in paraffin or a carrot, and the section mounted in Dammar. Sections of the uterus and ovaries are best mounted in glycerine or Dammar. Sections of lung maybe made by embedding in gum or by freezing, and mounted in Farrant's solution.
Every slide should be of uniform size, and labeled. The usual size is 3×1 inches, and should be of a good quality of glass, free from scratches or air holes. They may be labeled either by writing with a diamond, or a small piece of paper affixed to one end, on which is written what is required.