CHAPTER VI EXAMINATION OF MARINE OBJECTS DISSECTION

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An enthusiastic observer of nature will learn much concerning the structure of natural objects with the unaided eye, but there are times when he will desire some kind of magnifier to reveal more perfectly the structure of minute parts, or to enable him to observe the small creatures that are invisible to the naked eye. Further, one may learn many interesting and instructive facts relating to animal and plant life by cutting sections for close examination, or by making such simple dissections as will enable one to observe the more salient features of internal structure; we therefore propose in the present chapter to make a few remarks and suggestions regarding work of this kind.

A pocket magnifier is of great value to the young naturalist, both for the inspection of natural objects while engaged in out-door work, and for the subsequent examination of the specimens collected for study. It is often necessary to enable one to identify and classify small animals and plants, and will be in constant demand for the purpose of studying the less conspicuous external features. Such an instrument should be regarded as an essential companion of the naturalist, and should accompany him on every ramble.

There are several different forms of pocket lenses, but for general work there is, perhaps, nothing more convenient and serviceable than the ‘triplet’ magnifier. It is a combination of three lenses, enclosed in a pocket case, and so arranged that they may be used separately or in combination, thus supplying a variety of powers. The three lenses of the triplet are themselves of different magnifying powers, and these powers may be increased by combining two or all of them.

For work at home a ‘dissecting microscope’ is very useful. This consists of a magnifying lens, mounted on a support over a surface on which small objects may be examined and dissected, the height of the lens being, of course, adjusted according to its focal distance. Lenses ready mounted on adjustable stands may be purchased for this purpose, but no one ought to experience much difficulty in designing and constructing some simple stand that will give every satisfaction.

The arrangement just described is, of course, suitable for the dissection of only small objects, and these are placed on a material adapted to the nature of the work to be done. Thus it is sometimes convenient to place the object to be examined on a small sheet of cork, in order that it may be secured by means of pins while the dissection proceeds, while at other times it is essential that it be laid on a hard and unyielding surface, such as that of a slip of glass. But whatever be the nature of the substance on which the dissection is made, its colour may be regulated according to that of the object. If, for example, we are dissecting a small white flower on a piece of cork, we should naturally blacken the cork, or cover it with a piece of dead black paper; or, if we are to dissect a small, light-coloured object on a glass surface, we lay the glass on black paper.

Fig. 45—The Triplet Magnifier

The advantage of dissecting objects under water does not seem to be generally appreciated by beginners, who often allow their specimens to become dry and shrivelled, almost beyond recognition, during the progress of their examination. This mode of dissection is certainly not necessary with all objects, but may be generally recommended for soft and succulent vegetable structures, as well as for almost all animal dissections.

This being the case, arrangements should certainly be made to provide a miniature dissecting trough as an accessory to the dissecting microscope, and the following instructions will enable the reader to construct a highly satisfactory and inexpensive one:—

Procure the flat lid of a cylindrical tin box, or the lid of a glass or porcelain pomade pot, such lid to be about two inches in diameter and about half an inch in depth. Cement the flat side of this lid to a small slab of hard wood, or to a square piece of sheet lead, by means of acetic glue—ordinary glue or gelatine dissolved in glacial acetic acid—to give it the necessary steadiness during the dissection. When the cement is quite hard, pour into the lid some melted paraffin (paraffin wax) which has been blackened by the admixture of a small quantity of lamp-black in the form of a fine powder. The paraffin should be melted by putting it into a beaker or wide-mouthed bottle, and standing it in hot water, and the lamp-black should be added, with stirring, as soon as it is entirely liquefied. The quantity of the mixture used must be sufficient to half fill the lid, thus leaving a space to contain water to the depth of about a quarter of an inch. The blackened wax provides a good background on which to work, and provides a hold for pins when these are necessary in order to fix the object under examination.

Fig. 46.—A small Dissecting Trough

The complete trough is represented in fig. 46; and will be found to answer its purpose admirably, except that it occasionally displays one fault, but one that is easily remedied. The wax contracts on cooling, and may, therefore, detach itself from the trough; and, being lighter than water, will float instead of remaining submerged. This may be prevented by securing the disc of wax in its place by means of a ring of brass wire, or by weighting the wax with two or three small pieces of lead pushed down into it while it is yet soft.

With such a dissecting microscope and trough as we have described one may do a great deal of exceedingly useful work, both hands being quite free to manipulate the object under examination.

The dissection may be conducted with the aid of a small scalpel or other very sharp knife, the parts being arranged or adjusted by means of a needle, mounted in a handle, and held in the left hand. Sometimes, however, the object to be dissected is so minute that even a small scalpel is too large for the purpose, and in such cases nothing is better than little dissecting instruments made by mounting large sewing needles in suitable handles, and then grinding down the points of the needles on two opposite sides, on a hone, so as to produce little pointed, two-edged blades. Bent needles are often useful, too, and these may be prepared by heating the points to redness in a gas-flame, bending them as desired while hot, and then hardening them by suddenly thrusting them, at a red heat, into cold water.

The compound microscope will often prove useful for the examination of very minute objects, as well as for the study of the structure of the principal tissues of the larger species; but since detailed instructions for the management of the microscope, and for the preparation of objects for microscopic examination would occupy much more space than we can spare, we shall content ourselves with nothing more than a few general hints on this portion of the young naturalist’s work, dealing more particularly with those points which commonly present difficulties to the amateur.

If it is desired to examine some minute living object, such as a protozoon, place the object in a drop of the water in which it lived just in the middle of a clean glass slip, and cover it with a cover-glass. The quantity of water should be just sufficient to fill the space between the two glasses. If less than this has been used, a little more applied to the edge of the cover by means of a glass rod will immediately run in between the glasses; while if an excessive amount was employed, the surplus may be removed by the application of a strip of blotting paper. Place the glass slip on the stage of the microscope, and reflect light through it from the mirror below.

Examine it first with a low power; and, after having observed as much as possible of the creature’s movements and structure with this aid, repeat with a higher power. This rule applies not only to such small objects as we have now under consideration, but to all objects, and parts of them, in which minute details are to be observed.

Beginners with the microscope often find prolonged examination very tiring to the eyes, but this, we believe, would seldom be the case if right methods were followed. Both eyes should always be open, and the microscopist should train himself to use both eyes equally for the actual observation.

The higher the magnifying power used, the nearer must the objective (the lower combination of lenses) be brought to the object itself, and it is no uncommon thing for the amateur, in his attempts to focus his object, to lower the body of the microscope beyond its proper position, causing the objective to crush the object, break the thin cover-glass, and become wetted with the liquid, if any, in which the object was being examined. All this may be avoided by lowering the body of the microscope until it nearly touches the cover-glass before attempting to view the object through it, and then, with the eye above the object-glass, to gradually raise the body until the object is in focus.

Fig. 47.—Cell for small Living Objects

The top of the cover-glass should always be perfectly dry; and if by any chance the objective becomes wet it should be wiped perfectly dry with a piece of old silk or with chamois leather. Also, if permanent mounting is attempted, and the preservative liquid is allowed to come in contact with the objective, such liquid must, of course, be washed off with some suitable solvent before any attempt is made to wipe the lens dry.

If the object under examination is of such dimensions that the cover-glass has a tendency to rock on it, or if it is a living object of such a size that it is unable to move freely in the exceedingly thin film of water between the cover and the slip, it should be placed in a cell. The cell may be made by cementing a ring of glass or vulcanite to the middle of a slip, or it may be a little circular cavity prepared in the slip itself. In either case the cell must be quite full of water before the cover-glass is applied, so that no air-bubbles are included.

Hitherto we have spoken only of mounting small objects in water, and this is advisable when the object is moist, whether it be animal or vegetable, alive or dead. But dry objects may be examined in the dry state, in which case they need not be covered. If they are composed of transparent material they are to be dealt with in the manner recommended before, as far as the management of the light is considered; that is, a moderately strong light is sent through them by the reflector below the stage; but opaque objects are best examined on a dead black ground, the light being directed on to them by means of a condensing lens placed between them and the source of light.

A collector who has done only a few days’ work on the sea shore will probably find himself the possessor of a host of interesting objects that will afford much pleasure and instruction when placed under the microscope—objects, many of which have been somewhat hastily deposited in a bottle of spirit or other preservative for study in his future leisure moments. These objects, if small, may be examined as above described, simply placing them under a cover-glass, or in a cell, with a clear drop of the same liquid in which they have been kept.

The general characters of the larger objects may also be observed by means of some kind of hand lens, but even these are generally best examined under water or other suitable liquid.

A great deal may be learnt of natural objects by preparing very thin sections for microscopic examination; and although special works should be consulted if one desires to become proficient in the different methods of cutting and preparing such sections, yet a great amount of good work may be done with the aid of a sharp razor, manipulated with nothing more than ordinary skill.

Some objects, especially certain of those of the vegetable world, are of such a nature that suitable sections may be cut, either from the fresh or preserved specimen, without any preliminary preparation. All that is required is to hold the object firmly between the finger and thumb of the left hand, previously securing it in some kind of holder if necessary, and pare off the thinnest possible slices with a horizontal movement of the razor, both razor and object being kept very wet during the process. As the sections are cut they may be allowed to drop into a shallow vessel of water; and, the thinnest then selected for examination in water as previously described.

Other objects are so soft that the cutting of sections becomes impossible without previously hardening them. Methylated spirit is a good hardening reagent, and many of the soft structures that have been preserved in this fluid, especially if it has been used undiluted, will be found sufficiently hard for cutting thin sections. Among the other hardening reagents used by microscopists may be mentioned a solution of chromic acid—one part by weight of the solid acid dissolved in from one hundred to two hundred parts of water, and a solution of bichromate of potash—one part of the bichromate to about forty parts of water. In either case the hardening of the object takes place slowly, and it should be examined from day to day until the necessary consistence has been obtained.

The structures of many soft animals can never be satisfactorily hardened for section-cutting by either of the above reagents, and thus it becomes necessary either to freeze or to imbed them. In the former case the object is first soaked in gum water—a thin solution of gum arabic—and then frozen by an ether spray or by a mixture of ice and salt. The sections should be cut with a razor just as the object is beginning to thaw, and they may then be examined under a cover-glass, in a drop of the gum water.

The other method is conducted as follows:—The soft object is first soaked in absolute alcohol to extract all the water it contains, and is then transferred to paraffin that has been heated just to its melting-point by standing it in warm water. After the object is thoroughly permeated with the paraffin, the whole is cooled quickly by immersion in cold water. Sections are now cut, the paraffin being sliced away with the substance it contains. These sections are placed in warm turpentine, where they are allowed to remain until the whole of the wax has dissolved, and they may then be mounted in a drop of turpentine, and covered with a cover-glass.

We have given brief instructions for temporary mounting only, but most amateur microscopists would undoubtedly prefer mounting their objects permanently, so that they may be set aside for study at any future period. Hence we append a few directions to this end, advising the reader, however, to consult a work dealing especially with this subject if he desires to become proficient in the preparation of microscopic slides.

Moist objects, including those which have been preserved in dilute spirit, may be soaked in water, then transferred direct to the glass slip, and covered with a drop of glycerine. Any excess of the glycerine should then be absorbed from around the cover-glass by means of a strip of blotting-paper, and the edge of the cover cemented by gold size applied with a small camel-hair brush.

Glycerine jelly is also a valuable mountant for permanent work. When this is used the object should first be soaked in glycerine, and then in the melted jelly. It is then transferred to a drop of melted jelly which has been placed on a warm slide, and covered as before. The jelly soon solidifies, so that a ring of cement is not absolutely necessary, though it is advisable, as a rule, to cement the cover-glass all round with gold size or black varnish.

Sections cut while frozen are best mounted in glycerine, to which they may be transferred direct.

Canada balsam is one of the best media for permanent mounting; and, as it becomes very hard after a time, it serves the purposes of both preservative and cement. When this is used the object must be entirely freed from water by soaking it in absolute alcohol. It is then put into turpentine for a minute or two, transferred to a warm slide, and covered with a drop of the prepared balsam. Sections that have been imbedded in paraffin may be mounted in this way, the turpentine acting as a solvent for the paraffin in which it was cut.

Although the compound microscope is absolutely necessary for the study of the minutest forms of life and of the minute structure of the various tissues of larger beings, yet the young naturalist will find that a vast amount of good work may be done without its aid. Thus the general structure of the larger species may be made out by means of simple dissections requiring no extraordinary skill on the part of the worker, and with appliances that may be obtained at a low cost. Certain of the marine animals, however, require special treatment that can hardly be described in a short chapter devoted to general instructions only, but hints with regards to these will be given in future chapters in which the animals referred to are described.

The appliances referred to above include nothing more than a simple form of dissecting trough, a few dissecting instruments, and one or two minor accessories that may always be found at hand as required.

The dissection of animals is always best performed under water, for by this method the object examined may not only be kept clean as the work proceeds, but the parts, having a tendency to float, readily separate from one another and therefore become more distinctly visible when submerged.

Fig. 48.—Sheet of Cork on thin Sheet Lead

A very convenient form of trough may be made by taking any kind of rectangular, flat-bottomed dish, one made of zinc being, perhaps, the best of all, and covering the bottom with a slab of good cork carpet which has been weighted with sufficient lead to prevent it from floating. Or, instead of cork carpet, a sheet of cork may be used. In either case, a piece of thin sheet lead, a little larger than the slab, should be cut, the corners of which are then snipped off as shown in fig. 48, and the edges finally turned over as represented in the next illustration. The size of the trough must be regulated according to the nature of the work to be done, but one measuring ten inches long, seven wide, and two inches deep will answer most purposes.

Fig. 49.—Weighted Cork for Dissecting Trough

The object to be dissected is placed in the trough, secured in position by means of a few ordinary pins, and then completely covered with water.

We need hardly impress upon the reader the great importance of thoroughly examining all external characters—all those structures that are visible without actual dissection—before attempting to remove anything; and we have already insisted on the importance of carefully examining all creatures while alive before anything else is done. The value of this latter stipulation can hardly be overestimated, for in many instances it is almost impossible to detect the use of an organ unless it has been observed in action; and the enthusiastic student will go even further than this, for he will make it an invariable rule to sketch everything he sees, and to make full notes on all his observations.

When pins are used to fix the object under examination—and it is generally essential that the object be fixed—their heads should be turned outwards; for then the object will not slip from its position, nor will the pins tend to get in the way of the work.

Some objects are of such a nature that they are not easily secured by means of pins, and yet require to be fixed in some way or other. Thus, one may desire to examine the structure and appendages of a prawn or small crab, or to investigate the nature of a chiton. In such instances as these it is a good plan to make a cake of paraffin wax of suitable size by pouring the melted substance into a mould, then secure the object in proper position in the wax while still fluid, and pin the latter to the cork of the dissecting trough.

It is often necessary to trace the courses of internal passages that open on the surface of the body, or of tubes that are revealed during the progress of dissection, and this may be done by means of a little instrument called a seeker. It is simply a blunted needle, bent into a large angle, and mounted in a handle; or, it may consist of nothing but a moderately long and stiff bristle, rendered blunt at one end by tipping it with melted sealing wax. This is not always sufficient, however, for it frequently happens that certain tubes and passages in animal forms are disposed in such a complicated manner that it is impossible to send even the most flexible seeker through them. For instance, suppose one desires to trace the course of the digestive tube of some large bivalve mollusc with its many reflections, the seeker is useless except that it will penetrate to the first sharp bend. The arrangement of such a tube must be traced by dissecting along its course, but this may be aided considerably by first filling it with some coloured substance to enable its direction to be more easily followed. In fact, the injection of some brightly coloured fluid, forced through the tube by means of a fine-nozzled glass syringe will often enable the course of such a tube to be seen without any dissection at all, the colour of the fluid used being detected through the semi-transparent tissues surrounding it. A mixture of Berlin blue and water, or a mixture of plaster of Paris and water coloured with carmine is well adapted to this purpose; and if the latter is employed it may be allowed to set, and thus produce a permanent cast from the tube that is being dissected. Perhaps it should be mentioned that if either of the injection mixtures be used for this purpose it must be previously strained through muslin, and that, in the case of the plaster, the mixing and straining should occupy as little time as possible, or it may begin to set before the injection has been completed.

A very considerable insight into the structure of animals may be frequently obtained by cutting sections through the body with all its organs in situ, but, generally speaking, they are too soft to allow of this without danger of the displacement of those very parts, the relations of which we desire to determine. To avoid this the body should be previously hardened by a somewhat prolonged soaking in methylated spirit, or in a solution of chromic acid prepared as before directed. Then, with the aid of a good razor, very interesting sections may be prepared with the greatest of ease, and the true relations of the various organs throughout the body may be exactly determined by cutting a succession of slices, not necessarily very thin, from end to end, or, transversely, from side to side.

Even those crustaceans that are protected by a hard, calcareous exo-skeleton, and the molluscs that cannot be removed from their stony shells without injury to their soft structures, may be studied in the manner just described, and this may be done by first soaking them in dilute hydrochloric acid, renewed as often as may be necessary, until all the mineral matter has been dissolved completely, and then hardening the softer tissues in one of the reagents mentioned above. Hydrochloric acid may also be used to dissolve the calcareous shells of foraminifers, the vegetable corallines, and other small forms of life, previous to microscopic examination of the soft parts.


                                                                                                                                                                                                                                                                                                           

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