If the preceding dissections have been done, the crystalline lens will already have received some notice. To study the lens properly one should use an eye that has not been hardened and also an eye or the lens of an eye that has been in a 5 per cent. solution of formaldehyde for about two weeks.
The lens in the unhardened eye will prove too friable to permit much handling. The dissection should be made, however, in order to give opportunity to notice the crystalline clearness of the lens substance, its great magnifying power, its attachments, its capsule, etc. For this purpose it is necessary to proceed only as in the dissection for the “hyaloid membrane, etc.” That is, use an eye that has been kept in a cool place for several days, and then open it, and remove hyaloid, vitreous, and lens intact, as in the first dissection taken up in this book. To examine the specimen in detail, turn it so the lens will be uppermost. (Fig. 18.)
To remove the lens it is necessary to separate the suspensory ligament, using for this purpose the small-pointed scissors. The capsule may be removed by picking it up on the periphery of the lens, and stripping it off. It will peel off about the same way that the outer skin of a bean or pea does.
The tri-radiate lines on the posterior and the anterior surfaces of the lens will not be as clearly discernible as in the lens coming from the hardened eye. Close inspection and the use of a hand lens will help bring them out more clearly.
Now, with the point of the scalpel try to separate the outer layers (cortex) from the harder inner layers (nucleus). This will not prove very successful but is suggested for the purpose of comparison when the same thing is done to the hardened lens.
It will be found that the lens after having been in the formaldehyde solution is no longer crystal like, but more or less translucent. When viewed from either the anterior side or the posterior side, the tri-radiate lines on each surface will be seen to begin at the poles of the lens and radiate outward toward the lens equator. Holding the lens up to a strong light will show that though the lines on either surface form angles of 120 degrees, the angles formed by the lines on one side with the lines on the other side are 60 degrees. On the anterior surface of the lens the vertical line extends upward from the pole; on the posterior surface downward from the pole.
To study the laminated structure of the lens, it is best to boil the lens. The best way to do that is to drop the lens from either a hardened or unhardened eye into boiling water. Let it boil in the water for about two and a half to three minutes. Longer than that time will cause the lens to be put out of shape, and make it so fragile that it can no longer be handled without having it fall apart. If the lens comes from an unhardened eye it might be best to boil it not more than about two minutes.
Insert the point of the scalpel carefully at one of the poles, and lift gently in the direction of one of the radiating lines. This will tend to raise one of the concentric layers, which can be easily peeled off. Repeat this in the direction of the other two radiating lines. Examining, with a hand lens, the exposed surfaces and the layers, as they are taken off, will show the arrangement of the lens fibres, and will also show plainly their directions. (Fig. 19.) To get another view of the onion-like layers of the lens, cut through it with a safety-razor blade, either longitudinally or equatorially. (Fig. 20.) The better way is to have enough lenses to make one of each kind. Never try to work with only one piece of material. If the lens is first stained with chromic acid the layers may be seen better, or, a simpler way is to drop the lens, before cutting it in two, into a carmine solution; red ink slightly diluted, will do.
A lens that has been boiled and partly dissected may be placed in a 5 per cent. formaldehyde solution, and kept indefinitely. The lens fibres, concentric layers, and lens laminae in such a specimen will always be interesting.
A lens that has lost its transparency because of hardening in formaldehyde or boiling may be made clear and nearly transparent again in the following way: First: Place the lens in a 50 per cent. alcohol for several hours. Second: Remove the lens, and let it drain on a piece of blotting-paper; then place it in a 75 per cent. alcohol. Third: Remove the lens, as before, then place it in an 85 per cent. alcohol. The lens may be left in this alcohol from ten to twelve hours, after which length of time it should be removed and drained. Fourth: Place the lens in an absolute alcohol, and leave it there for ten or twelve hours. Several hours longer will not injure the lens, nor interfere with the success of the work. Fifth: Remove the lens from the absolute alcohol. Place it upon a piece of blotting-paper, moving it to another place on the blotting-paper whenever the paper around the lens seems to have taken up as much moisture as it can hold. Be sure that the lens has given up nearly all, if not all, moisture. “Running through the alcohols,” as this process is called, is for the purpose of dehydrating the tissue. It will be on the side of safety to let the lens lie exposed on the blotting-paper for an hour. Sometimes, if the capsule has not been removed, a small quantity of alcohol will remain between the lens and the inner surface of the capsule. This must be removed. It may be done by either puncturing the capsule with a pin or needle, and squeezing out the fluid, or by removing the capsule entirely. The latter is preferable.
Now drop the lens into xylol. Benzine will answer, though it will not produce quite so clear a lens as the xylol does. At the end of 24 or 36 hours the softer cortex will show quite clear, while the harder nucleus will be still cloudy. At the end of a week the whole lens, if it is a small one--pig, calf, sheep--will have become quite clear and transparent; if from a beef eye it will take longer. It sometimes takes nearly two weeks. In the case of a boiled lens it will take much longer to clear; it may take a month.
Cedar oil may also be used for the purpose of clarifying or “clearing” the lens. Harden in the usual way, run through the alcohols, and then place in cedar oil. The oil, however, will stain the lens a yellowish brown, and the lens will not be as transparent and clear as when xylol is used.
A. Lens hardened in formaldehyde.
B. Lens hardened in formaldehyde, run through the alcohols, and cleared in xylol.
C. Lens hardened in formaldehyde, run through the alcohols, and cleared in cedar oil.
D. Boiled lens.]
The longer a lens is left in either of these two clarifying fluids the harder and smaller it will become. At the end of a month or six weeks the lens will have become so hard that it can no longer be cut through with a knife. If it is desired to halve it, a scroll saw will be found to be the best thing to use for this purpose. (Fig. 21.)
“Anatomy and Physiology of the Eye,” Brown & Zoethout.
THE CHOROID
Select an eye that has had a long part of the optic nerve left on it and place it into a 5 per cent. solution of formaldehyde. Leave it in that solution for from two to three weeks. That period of time in the fluid will be sufficient to permit the choroid to become sufficiently toughened and hardened. Leaving it in the solution longer than that length of time will not injure the eye in any way.
After having removed the eye from the formaldehyde, wash it for a few moments in running water. This will remove the preserving and hardening fluid from the surface, and will save the hands a little from the effects of the fluid. Next remove all the muscles and fatty tissues from the sclerotic. After that has been done, puncture the cornea with the pointed jaw of the scissors about 2 mm. from the corneo-scleral junction. (Fig. 22.) Then proceed to cut the cornea away, being careful not to lacerate the choroid or the iris. (Fig. 23.) The escaping aqueous humor will flow over the eye and make it very slippery, and, therefore, difficult to hold. Dip the eye in water, wash it, and then take it out and thoroughly dry it with a cloth. This procedure is absolutely necessary, and, if omitted, will surely result in the dropping of the eye about the time the work on the specimen is nearly finished. Insert the scalpel between the peripheral edge of the exposed iris and what is left of the cornea. With the back edge of the scalpel, gently loosen the choroid from the inner side of the corneo-scleral junction to which part it is not securely attached. (Fig. 24.) This requires only ordinary care, and but little skill other than that necessary to always keep the scalpel close to the inner surface of the sclerotic. When the choroid-iris edge has been detached from the inner side of the corneo-scleral junction, the weight of the contents of the second tunic will cause it to sag and give opportunity to easily separate, with the back edge of the scalpel, the choroid from the sclerotic for about a distance of from 8 to 10 mm.
It has been the method in the past to force water through a blowpipe between the sclerotic and the choroid, in order to separate the attachments. It has also been the method to work under water when wishing to expose or isolate either the choroid or the retina. It is unnecessary to do either of these two things.
When the sclerotic has been loosened from the choroid for about 10 mm. back from its cut edge around the eye, carefully cut the loosened part away. (Fig. 25.) Then loosen the choroid as far back as to within 1 cm. of the optic nerve. Cut the separated sclerotic away. It will be well to state here that during this dissection the specimen should not be lifted from the table. Keep the eye resting on the table all the time, and never lift it by holding it suspended from the optic nerve. Loosening the choroid from the sclerotic up to this point is a very easy matter; ordinary precaution is all that is necessary to prevent puncturing the choroid with the scalpel, just be sure to remember to keep the point of the scalpel close to the sclerotic.
To remove the remaining part of the first coat is a little more difficult, and needs a little more care. Hold the optic nerve in the left hand, and pull it so that the sclerotic will pull away from the choroid. Then, using the cutting edge of the scalpel, scrape the choroid loose from the sclerotic close up to the entrance of the optic nerve. (Fig. 26.) Do not separate the optic nerve from the choroid. Cut away the remainder of the sclerotic close up to the optic nerve and the choroid will be free. (Fig. 27.)
To get a perfect specimen and completely isolated choroid, it must be emptied of its contents. Insert the scalpel between the lens and the iris, force it on through, and in such a manner as to keep the scalpel close to the ciliary processes. (Fig. 28.) Cut the vitreous around the processes. Push the scalpel further into the vitreous, and cut out the central part of it, just as one would cut out the core of an apple. (Fig. 29.) Remove the scalpel, pick out the lens and the cut centre of the vitreous with the broad-point tweezers, holding the choroid a trifle suspended by the optic nerve. The remaining part of the vitreous may be broken down by cutting with the scalpel, and by squeezing and crushing with the fingers of both hands. (Fig. 30.) The choroid will be tough enough to stand this treatment provided the pupil is left clear and open to prevent inter-choroidal pressure. After the vitreous has been removed the choroid will be left in a greatly collapsed condition. Dropping it into water and letting it fill up will make it resume its original shape immediately. The retina does not always come out with the vitreous. In such a case, the tweezers may be used to pick out the retina when the choroid is in a collapsed condition.
This specimen will show the vena vorticosa, the ciliary nerves, and their way of ramifying, and the long ciliary arteries, which run opposite each other and which may be recognized by their rather colorless, tubular appearance. The evacuated choroid makes an excellent specimen and one easily examined. Place it in a 3 per cent. solution of formaldehyde, and then examine with a skiascope, an ophthalmoscope, or by “oblique illumination.”
This dissection is wholly original, and may be done in about five minutes. The old technique for doing it required at least an hour of time with the possibility of procuring one perfect specimen in every six or seven. The technique as given here will make it possible to do the work in not longer than five or six minutes for the beginner, and about four minutes for the expert.
THE RETINA
Technique of Eye Dissections · The Wunder Library — complete classics, free to read, with narration.