Formalin is an excellent preservative fluid; one part to 20,000 is sufficient to prevent fermentation. For the preservation of vegetable sections, a one per cent. solution is required; even the fresh appearance of vegetable structures is preserved for some time when immersed in it. In the nutrient gelatine for biological specimens, if used early, will arrest the liquefaction of the gelatine by bacteria. For hardening it saves time, and is even better than alcohol, chromic acid, pot. bich., and many others. It does not cause shrinkage of the cells. Tissue ½ to ¾ inch thick hardens in twenty-four hours in pure formalin; five to ten per cent. is best for loose tissue. In another method, by which time can be saved, instead of placing the specimen in the formalin and afterwards in mucilage, prior to cutting sections, make the mucilage with two per cent. (or stronger) formalin water, and it will then answer both purposes at the same time.
Preparing, Mounting, Cementing and Collecting Objects.
Various materials are required for preparing and mounting microscopic objects, as slips of glass, patent flatted plate measuring 3 × 1 inch, thin glass covers, glass cells, preservative media, varnishes, cements, a glazier’s diamond, and a Shadbolt’s turn-table.
The glass slides and covers, although sent out packed ready for use, should be immersed in an alkaline solution to ensure perfect freedom from any greasiness derived from touching by the fingers. Dr. Seller recommends a particular solution for this purpose. (See Formulæ, Appendix.)
Varnishes and cements must be selected with care, as these are not only expected to adhere firmly to the glass slide, but also to resist the action of the preservative fluid in which the specimen may be mounted. Among the numerous preparations employed, I may enumerate Canada balsam, gum dammar, Venice turpentine, Japanners’ gold size, used for closing up cells, asphalte varnish, Brunswick black, shellac, glue and honey, Hollis’ liquid glue, and marine glue. To give a finish to the mounted specimen, coloured varnishes are sometimes resorted to. A red varnish of sealing-wax is made by digesting powdered sealing-wax in strong alcohol. Filter, and place the solution in a dish, and evaporate by means of a sand bath to reduce it to a proper consistency. This is said to resist the action of cedar oil. For white, zinc, cement is the best. This is made of benzole, gum dammar, oxide of zinc, and turpentine. Cole gives another formula, but either of these may be obtained of Squire, who supplies every kind of staining and mounting material.
Fig. 262.—Walmsley’s Cell-making Turn-table.
Cells for Mounting.—The minuter forms of life should be mounted in thin cells, which may be readily made with Japanners’ gold size, dammar or asphalte, and a Shadbolt or Walmsley’s turntable. The glass slide being placed under the metal springs in such a manner that its two ends shall be equi-distant from the centre (a guide to the position is afforded by the circles traced out on the brass), take a camel’s hair pencil and dip it into the Japanner’s gold-size, holding it firmly between the finger and thumb, and set the wheel in motion, when a perfect circle will be formed; put it aside to dry, or place it in the warm chamber to harden. To cut cover-glasses place a sheet of thin glass under the brass springs, and substitute for the pencil a cutting diamond. A cutting diamond is not only useful to the microscopist for the above purpose, but also for writing the names of mounted objects on one end of the slide.
It will be found convenient to make a number of such cells, and keep a stock ready for use. There are many objects whose structure is very transparent. These should be mounted dry. Scales from the wings of butterflies and moths, of the podura and lepisma, and some of the diatomaceæ are of this class. All that is necessary in preparing objects for dry mounting is to take care that they are free from extraneous matter, and fix them permanently in the position in which their structure will show to the best advantage.
For mounting specimens of greater thickness it is desirable to use deeper cells. It will then be found convenient to make a second or a third application of the gold-size, allowing sufficient time between applications for the varnish to dry. Cells of a still deeper kind are made up by cementing rings of glass or metal to the glass-slides with marine glue or Brunswick black. The latter will be rendered more durable by mixing in a small quantity of indiarubber varnish (made by dissolving small strips of caoutchouc in gas-tar). The process of mounting in glass-cells is similar to that employed in making varnish-cells, except that a somewhat larger quantity of cementing medium is required. Objects mounted in this way should be kept for a time in the horizontal position, and a little fresh varnish must be applied if the cement shows a tendency to crack. In mounting objects in balsam, care must be taken to have the specimen quite dry before transferring it to turpentine. Objects mounted in cells should become perfectly saturated with the mounting fluid before being finally cemented down.