Demonstration that the red globule is a real cellule having microzymas for anatomical elements.

Molecular granulations of the globules of fowl's blood.

Molecular granulations of the globules of the duck's blood.

An exact knowledge of the physical constitution and of the anatomical structure of the red globule is of great im­portance to the scope of this work. Is or is not the red globule of the blood a cellular anatomic element, constituted by an envelope with its content; or is it a kind of naked anatomical element, as was said of the milk globules? An alternative, necessary to be resolved, to obtain a clear idea of the role of the red globule!

Prevost and Dumas1 admitted that there was an envelope to the red globule and, later, Henle,2 by exact observations, demonstrated the reality of the existence of this envelope. In 1856, Kuss taught us in his course on physiology, at Strasbourg, that "the blood globules are not bladders, but compact organs, solid in all their parts, the least aqueous of all the organs of the body." More than twenty years afterwards, M. Frey, interpreting the general opinion, said: "To sum up, the globule maybe regarded as a mass of gelatinous substance saturated with water."3 And he added: "In spite of the difficulties of observation and the uncertainty of such examinations, some authors have pronounced themselves, during these later years, in favor of the existence of a cellular membrane."

1.  "Ann: de Chimie," Vol. XVIII. p. 280 (1821). and Vol. XXIII, pp. 53 and 90 (1823).
2.  "Anatomie generale," Vol. I, p. 459. Jourdan's trans. (into French.)
3. Frey, loc. cit., p. 123.

Dumas, as we shall see, not only considered them as having a cellular constitution, but as individual living beings, saying that the deprivation of oxygen was fatal to them. Such was not the physiologists' point of view; and I cannot give a better proof of this than the following:

I had compared the scintillating corpuscle of the pebrine to a cellule already acknowledged to be alive, such as that of the globule of yeast. M. Pasteur asserted that this was an error and, referring to it, made the following statement: "My present opinion is that these corpuscles are neither animals nor plants. * * * From the point of view of a methodical classification they should be placed rather alongside of globules of pus or blood globules, or still better, of grains of starch rather than alongside of infusoria and moulds;"1 and later: the corpuscle "is a production which is neither animal nor vegetable and incapable of reproduction, and it must be placed in the category of those bodies, regular in form, which physiology has for some years distinguished by the name of organites, such as globules of blood, globules of pus, etc."2

1. C. R,Vol. LXI.p. 511.
2.  lbid, Vol. LXIII, p. 134.

It is evident that that which is neither plant nor animal has neither organized structure nor life; has no content in an envelope. But that was the state of science, and that which M. Pasteur believed regarding the blood globule and regarding pus, was believed of every other anatomical element, for instance, of the spermatozoids, comprised in the "etc." of M. Pasteur, and which he compared to grains of starch, under the name of organites, i.e., simulacra of organs. If I insist particularly on this savant's mode of viewing these things it is because he has specially occupied himself about the blood and thence about what happens to its globules during its spontaneous changes.

It is no idle question to inquire whether the blood glob­ule is naked or covered, furnished with an envelope separate from its mass. The property of a living body as to whose organization there is no dispute, whether it relates to an animal or to a globule of yeast, is to be limited in its form by a continuous enveloping membrane (distinct from its interior medium), which lakes the name of tegument, a word of which the etymological meaning is clear; the enveloping membrane of the cellule is then the same as the tegument. In the organism the interior organs are individualised by their own tegument. Among bodies which can only realize the conditions of their existence in water or in aqueous media, the insoluble tegument, endowed besides with special osmotic properties, protects the content, or the interior medium, against being dissolved or from other like change. Well! the blood globule like beer-yeast, is individualized by its tegument: hence it is an organ and not an organite.

That which has given rise to doubt as to the existence of a tegument to the red globule is that the blood steeped in water seems to be altogether dissolved in it: the globules disappear so completely that under the microscope not a vestige of them can be discovered. Those who, like Dumas, admitted an envelope, thought that it was broken and thus explained why the weight of the isolated fibrin of the clot was greater than that obtained by whipping. But, as we shall presently see, that apparent solution of the globules is only the osmotic issuance of the interior part, soluble in water, across the envelope: the cellular tegument remaining entire is invisible under the microscope only because its refracting index is the same as that of the ambient liquid.

Demonstration that the red globule is a real cellule having microzymas for anatomical elements.

The mode of demonstrating the unbroken tegument of the red globule of the blood, steeped in water, consists in rendering its refraction different from that of the liquid which results from this mixture. To do this the blood, defibrinated or not, is to be mixed with an equal volume of a solution of about 15 per cent, of creosoted soluble fecula; at the end of 24 hours one can see that the globules resist the contact with the water much better, the envelope being clearly visible. The experiment is most striking with the blood of the duck; in one of them, which had lasted three weeks, successive washings with a solution of soluble fecula and with water removed all the coloring matter, leaving for residue colorless globules, where the nucleus could be seen rolling about in the water-logged globule; sometimes even the colorless tegument could be seen to wrap itself around the nucleus. The tegumentary vesicle, after these washings, has sometimes become so pale that it was only visible after an addition of an iodide paint which colors it yellow; but it is not colored by the ammoniac solution of carmin, nor by that of the picrocarminate.

I have also similarly experimented on the blood of a fowl, of a pigeon, of a frog, of a dog, of an ox, of a guinea-pig; in all cases the vesicle is seen entire, but among the elliptical globules having a nucleus those of the fowl deserve more attention. And it is to be noticed that, in experiments which take up a long time, the nucleus of the blood globules of a bird ends by being resolved into fine molecular granulations which can be seen in the colorless envelope which remains whole. In the emptied vesicles of the circular globules, nuclei are never to be seen.1

After the publication of these experiments Profs. J. Bechamp and E. Baltus described the process of tinting by which in all cases the unbroken cellular tegument may be distinguished, even in blood merely steeped in water.2

1.C. R., Vol. LXXXV. p. 712.
2. Ibid., p. 761 (1877).

But that is not all: if there is no cellule without envelope, neither is there one without microzymas. The blood globules are, in fact, no exception. On the other hand, as I have before mentioned, in certain experiments the decolored vesicles of the blood vessels of the duck, emptied of their coloring matter, contained their nucleus reduced to molecular granula­tions; on the other hand, I have stated that the deposit of molecular granulations formed in defibrinated blood, with two volumes of alcohol, 35-40 per cent, added, came from the destroyed globules of this blood. It is necessary then, by direct experiment, to put this beyond doubt, the doing of which has brought to light some particularly interesting observations.

To solve the problem of the existence of these molecular granulations in the globules of the blood, these latter were isolated from the defibrinated blood, separated from all trace of fibrin by filtration on fine cloth, and four times its volume of a saturated solution of sulphate of soda, or of an analogous salt, was added to it.

Molecular granulations of the globules of ox blood.

The globules being received upon a filter with the usual precautions are washed once more with sulphate of soda, and are then treated with alcohol of 35-40 per cent., which ought to dissolve the coloring matter; this was, in fact, done by a washing in this alcohol until discoloration was as complete as possible and finally with water; the filter retains the molecular granulations mixed with the remains of the cellular envelopes, possessing the appearance and properties of those obtained from defibrinated blood treated directly with alcohol.

The peculiarities already presented by the globules of the bloods of the duck and of the fowl induced me to repeat these experiments upon these bloods.

Molecular granulations of the globules of fowl's blood.

The globules separated from the defibrinated blood of the fowl by the saturated solution of sulphate of soda are received upon a filter and treated in the same manner as was ox-blood, with alcohol at 35-40 per cent., until deprived of their color; while the washing was made with alcohol, the colorless residue upon the filter seemed pulverulent, as in the case of the ox's globules; but by continuing the water washing the pulverulent mass was transformed into a mucous mass. Then, much surprised, I made this further experiment.

The defibrinated blood of another fowl was treated as usual by twice its volume of 35-40 per cent, alcohol. The pulverulent deposit of molecular granulations was obtained in the ordinary way; the deposit received upon a filter and washed with weak alcohol became white, remaining pulverulent; but so soon as water was added the matter assumed the mucous condition of the granulations of isolated elliptical blood globules.

The mucous mass sets free the oxygen from oxygenated water; it dissolves only with difficulty in hydrochloric acid at 2/1000.

As a consequence, from this it is clear that the albuminoid atmosphere of the microzymian molecular granulations of the blood globules of the fowl, which assume the mucous state in water, is formed of a different substance from that of the atmospheres of the blood globules of the ox. We have already seen that the fibrin of fowl's blood, yielded by whipping, is scarcely attacked by hydrochloric acid. The granulations of the globules of the same blood having become mucous have the same property.

Molecular granulations of the globules of the duck's blood.

The red globules of the duck, isolated from the defibrinated blood by sulphate of soda, are also treated on the filter by alcohol at 35-40 per cent. The most prolonged washing with this alcohol does not at all discolor the molecular granulations; much more abundant than in the fowl's blood they remain, until the end, colored of a brownish red. Is it, otherwise, with those of the deposit of the same defibrinated blood treated with alcohol?

The defibrinated blood of the duck treated at the moment bleeding, with two volumes of the same alcohol, rapidly gives a deposit much more abundant that does the defibrinated blood of the sheep. The deposit formed chiefly of the molecular granulations is not decolored by the washings with alcohol and with water, and remains of a brownish red.  Treated with very diluted hydrochloric acid it yields colored solutions.

The bloods of the fowl and the duck deserve a more complete study than, to my regret, I have been able to make.  For the results prove that the globules, elliptical in form and with a nucleus, differ not only by their forms from the circular globules, but further that they differ among themselves. These two facts, and those relating to the blood of the ox, prove that from this time forth we have to study, not only the blood, but the bloods, perhaps as much in the properties of their anatomical elements as in that of their albuminoid components, and especially of their haemoglobins.

However that may be, it remains proved that the blood globules in general are not only constituted upon the model of the perfect cellule, but that by a sojourn of the blood in a solution of fecula the refracting index of the tegument of the globules is modified without modifying its osmotic properties.

Further, it has been demonstrated that the blood globules themselves have, for anatomical elements, molecular granulations constituted like the haematic microzymian molecular granulations, as well the circular blood globules, as the elliptic, and that the molecular granulations of of one blood globule may differ from those of another. The physiology of the blood will be a great gainer by the study of of other special cases.1

1. This remark gives importance to the following: "The whole of the small family of Chameleons presents the singular exception that their blood globules are elliptical. But nothing like it has been found in other animals of the same class and nevertheless the bloods have been examined of more than 200 species chosen among all the natural subdivisions of the group, including even the marsupials and the monotrema, which, in certain respects, seem to establish the passage between the normal mammifera and the oviparous vertebrates. No adult bird is known whose blood globules are not elliptical. It is the same with reptiles, batrachians and ordinary fishes. Among cartilaginous fishes the lampreys, for example, the form of the globules is nearly circular." (Milne-Edwards, "Physiologic comparee. Vol. I, p. 48, etc., edition of 1857).
    The form, the external characters, among all living beings, are allied to the entirely of their other properties. Why should it not be the same with their blood globules and their other anatomical elements?
    [Milne-Edwards is correctly quoted by Prof. Bechamp, as above, but the statement of Milne-Edwards is not consistent with those of other authorities. All regard the chameleon as a reptile, and all say that the blood globules of all adult reptiles are elliptical.—Trans.]