The preparation & mounting of microscopic objects

Chapter III.

Chapter 131,877 wordsPublic domain

Sections of some exquisitely beautiful objects are cut with much less trouble than the above. The Orbitolite, for instance, may be prepared in this manner. Take the object and by pressure with the finger rub the side upon a flat and smooth sharpening stone with water until the portion is reached which it is wished to show. The strength of the object will easily allow this to be accomplished with ordinary care. This side may then be attached to the glass slide with heated balsam, as above described, and the object may then be gently rubbed down to the degree of thinness required to show it to the best advantage. After removing all disengaged matter from the object by washing and thoroughly drying, it may be mounted in balsam in the usual manner, when it is equally beautiful as a transparent or opaque object. From this it will be seen that in many instances where a smooth stone is found sufficient for the work (which is often the case when the section is mounted in balsam) the final process of polishing advised above may be dispensed with, as in the Orbitolite, Nummulite, &c., &c. It is quite necessary that the stones on which the objects are rubbed be _perfectly flat_, otherwise one side must be acted upon before the other, and it will be found impossible to attain anything like uniformity. Where it is not practicable to cut a section and the object is very thick, a coarse stone may be first used to reduce it and the smoother afterwards.

The consideration of the cutting of sections from shells would scarcely be deemed complete without some mention of what Dr. Carpenter terms the decalcifying process. Muriatic acid is diluted with twenty times its volume of water, and in this the shell is immersed. After a period, differing according to the thickness of the shell, the carbonate of lime will be dissolved away, and a peculiar membrane left, showing the structure of the shell very perfectly. This may be mounted dry, in balsam, or sometimes in liquid, according to the appearance of the object; but no rule can be given. The discretion of the student, however, will enable him to choose the most suitable method.

From some shells it is easy to divide thin plates, or “laminæ,” which require nothing but mounting in Canada balsam to show the texture very well. In working, however, with those which are “pearly,” it will be found that experience and patience are needed, as they are very brittle and peculiarly hard; but a little practice will overcome these difficulties.

Amongst the Echinodermata, which include the star-fishes, sea-hedgehogs, &c., there are many whose outer surface is covered with “spines,” or thin projections. Some of these are sharp and thorn-like, others blunt, longer or shorter, and, indeed, of endless variety. In many of these, when a section is made, rings are seen which have a common centre, with radiating supports, resembling sections of some of the woods. These are very beautiful objects, and methods of procuring them may now be considered. It is the best to cut as thin a section as can safely be got with the watch-spring saw first, when the smooth “sharpening stone” may be used to polish one side, which is easily accomplished with water only. When this is effected, it must be washed clean, and _thoroughly_ dried, and then may be united to the slide in the same manner as before recommended for the Orbitolite, &c. If it is ever necessary to displace it on account of inequalities, bubbles, or other remediable fault, this may be done by warming the slide; though too much heat must be avoided, otherwise fresh bubbles will certainly be produced. The covering with thin glass, balsam, &c., will present no difficulty to the student; but he must remember that the transparency is somewhat increased by this last operation.

_Corals_ are often treated in this way, in order to reveal their structure. Except, however, the student has had much practice, he will often find this a most difficult task, as many of them are exceedingly brittle. He will find the method before described equally applicable here, and should take both horizontal and vertical sections.

COAL.--This substance is one of the most interesting objects to the microscopist. It is, of course, of a vegetable origin; and though this is in many cases of such minute separate portions as to have lost all appearance of vegetation, yet it is very frequently met with in masses, bearing the form, even to the minute markings, of wood, in various directions. To see this and prepare it for microscopic research, a suitable piece of coal must be obtained; but in every case the cutting and preparation of these sections require great care and skill. Sometimes the coal is first made smooth on one side, fastened to the glass, reduced to the requisite degree of thinness, and finished in the method before described. This mode of treating it is sometimes, however, very tantalizing, as, at the last moment, when the section is about thin enough, it often breaks up, and so renders the trouble bestowed upon it fruitless. The dark colour and opacity of coal render an extraordinary thinness necessary, and so increase the liability to this accident.

Perhaps the best method which can be pursued is that recommended in the “Micrographic Dictionary,” which is as follows:--“The coal is macerated for about a week in a solution of carbonate of potash; at the end of that time it is possible to cut tolerably thin slices with a razor. These slices are then placed in a watch-glass with strong nitric acid, covered and gently heated; they soon turn brownish, then yellow, when the process must be arrested by dropping the whole into a saucer of cold water, else the coal would be dissolved. The slices thus treated appear of a darkish amber colour, very transparent, and exhibit the structure, when existing, most clearly. We have obtained longitudinal and transverse sections of coniferous wood from various coals in this way. The specimens are best preserved in _glycerine_ in cells; we find that spirit renders them opaque, and even Canada balsam has the same defect. Schulz states that he has brought out the cellulose reaction with iodine, in coal treated with nitric acid and chlorate of potash.”

_Cannel-coal_ is so close and firm in its structure as to be much used instead of jet in the manufacture of ornaments: it takes a beautiful polish, and consequently presents the student with none but ordinary difficulties in getting sections of it. Its formation is somewhat different from that of coal, sometimes showing the transition very clearly.

In _flint_ there are often found remains of sponges, shells, Diatomaceæ, &c.; but to show these well, sections must be cut and polished by the lathe and wheel of the lapidary, which the microscopic student seldom possesses. Thin chippings may, however, be made, which when steeped in turpentine and mounted in balsam, will frequently show these remains very well.

_Teeth_ are very interesting objects to all microscopists, more especially to those who give much study to them; as the class of animal may very frequently be known from one solitary remaining tooth. To examine them thoroughly, it is necessary to cut sections of them; but this is rather difficult to perform well, and needs some experience. Some instructions, however, will at least lessen these difficulties, and we will now endeavour to give them.

It is generally thought that Canada balsam injures the finer markings of these sections, consequently, they are almost invariably mounted _dry_. A thin piece is first cut from the tooth with the saw of watch-spring before mentioned, if possible; but should the substance be too hard for this, the wheel and lathe must be used with diamond dust. If this cannot be procured, there is no alternative but to rub down the whole substance as thin as practicable on some coarse stone or file. The surface will then be rough; but this may be much reduced by rubbing upon a flat sharpening stone with the finger, or a small piece of gutta-percha, upon the object to keep it in contact. The scratches may be much lessened by this, but not so thoroughly removed as microscopic examination requires in dry sections. It must, therefore, be polished with the putty-powder and dry strop, as recommended in the working of the shell-sections. The other side of the section of the tooth may then be rubbed down to the requisite thinness, and polished in the same manner, when the dust and other impurities must be removed by washing, after which the section must be carefully dried and mounted. Some of these sections are equally interesting as opaque or transparent objects.

The dentine of the teeth may be decalcified by submersion of the section in dilute muriatic acid; after drying and mounting in Canada balsam it presents a new and interesting appearance, showing the enamel fibres very beautifully when magnified about three hundred diameters. A friend tells me that after submersion of the _whole tooth_ in the acid he has been able to cut sections with a razor.

SECTIONS OF BONE.--With the aid of the microscope few fragmentary remains have proved so useful to the geologist and students of the fossil kingdom as these. From a single specimen many of our naturalists can tell with certainty to what _class_ of animal it has once belonged. To arrive at this point of knowledge much study is necessary, and sections of various kinds should be cut in such a manner as will best exhibit the peculiarities of formation. The methods of accomplishing this will now be considered. It may, however, be first mentioned that the chippings of some bones will be found useful now and then, as before stated with flint, though this is by no means a satisfactory way of proceeding. Sometimes the bones may be procured naturally so thin that they may be examined without any cutting; and only require mounting _dry_, or in _fluid_, as may be found the best.

When commencing operations we must provide the same apparatus as is needed in cutting sections of teeth, before described. A fine saw, like those used for cutting brass, &c.; two or three flat files of different degrees of coarseness; two flat “sharpening” stones; and a leather strop with putty-powder for polishing. As thin a section as possible should first be cut from the part required by the aid of the fine saw; and it is better when in this state to soak it for some short time in camphine, ether, or some other spirit to free it from all grease. With the aid of a file we may now reduce it almost to the necessary degree of thinness, and proceed as before recommended with teeth. The “sharpening” stone will remove all scratches and marks sufficiently to allow it to be examined with the microscope to see if it is ground thin enough; and if it is to be mounted _dry_ we must polish it with putty-powder and water upon the strop to as high a degree as possible, and having washed all remains of polishing powder, &c., from the section we must place it upon the slide and finish it as described in