Cited by W.B. Hardy, 'The Blood-corpuscles of the Crustacea, together with a suggestion as to the Origin of the Crustacean Fibrin-ferment (Part I)', Journal of Physiology 13 (1-2) (1892), pp. 165-190.
Description:

Explanation of Plate VII (figs. 1-25):

'Fig. 1. Living eosinophile cell fully loaded with granules. Oc. 4, Ob. D. [note: 'Oculars and Objectives referred to are those made by Zeiss'],  Cam. Luc.

Fig. 2. Nucleus surrounded by the remains of the cell substance of an explosive cell. Fresh preparation. Oc. 4, Ob. E. Cam. Luc.

Fig. 3. Group of eosinophile anid explosive cells after fixation with osmic vapour. The eosinophile cells show the characteristic vacuolate appearance (as seen in optical section), which results from the disappearance of the granules. Oc. 4, Ob. D. Cam. Luc.

Fig. 4. Eosinophile cell from which, as a result of its removal from the body, the greater number of the granules have been dissolved, leaving vacuoles. Fresh preparation.

Fig. 5. Eosinophile cell showing vacuolate endosarc. The blood of the animal from which this cell was taken contained numerous eosinophile cells, which, like the example figured, had discharged their granules while still within the circulation and as the result of some special stimulus. Fresh preparation. Oc. 4, Ob. D. Camu. Luc.

Fig. 6. Explosive cell undergoing division.

Figs. 7, 8, and 9. Stages in the explosion of an explosive cell studied by the aid of Iodine. Oc. 4, Ob. D. Cam. Luc.

Fig. 10. Eosinophile cell showing a stage in the colouration of the granules with indigo-carmine. Cam. Luc.

Fig. 11. Explosive cell. Osmic vapour. Oc. 4, Ob. E. Cam. Luc.

Fig. 12. Basophile cell from blood. Osmic vapour and Haematoxylin. Oc. 4, Ob. D. Cam. Luc.

Fig. 13. Basophile cell from blood to which a slight quantity of methylene blue has been added. Illuminated with ordinary gaslight. Cam. Luc.

Fig. 14. Basophile cell lying in a chamber of the basophile tissue. Fresh tissue treated with dilute methylene blue in normal salt solution. Ordinary gaslight, Oc. 4, Ob. D. Cam. Luc.

Fig. 15. Explosive cells treated with dilute iodine solution. The one to the left is the cell as first seen. The one to the right is the same cell as it appears about 15 m. after fixation. Oc. 4, Ob. D. Cam. Luc.

Fig. 16. "Phagocyte" from blood. Indian ink particles in heaps. Fresh preparation. Oc. 4, Ob. E. Cam. Luc.

Fig. 17. Vacuolate explosive cell from blood one hour after injection of Indian ink. Osmic vapour. Oc. 4, Ob. D. Cam. Luc.

Fig. 18. Explosive cell from blood one hour after injection of Indian ink. It contains little clusters of particles. Osmic vapour.

Fig. 19. Blood corpuscle of Daphnia as it appears in the living animal. Oc. 4, Oh. th, Zeiss. Cam. Luc. Owing to the very flat field of the objective used, only a few of the granules present are seen.

Fig. 20. Blood corpuscle of Daphnia containing a small heap of carmine grains. From living animal. Oc. 4, Ob. D. Cam. Luc.

Fig. 21. Blood corpuscle of Daphnica with granules stained by methylene blue. Ordinary gaslight. Oc. 4, Ob. E.

Fig. 22. Blood corpuiscle of Daphnia with small fat drops. Fronm living animal.

Fig. 23. Blood corpuscle of Daphnia with large fat drops. Oc. 4, Ob. D.

Fig. 24. Blood corpuscle of Dalphnia containing fat globules immediately after its attachment to abdominal fat tissue. Oc. 4, Ob. D. Cam. Luc.

Fig. 25. Basophile cell from blood of Astacus. Unstained.' (189-190)

 

Figs. 1 and 3 in text:

'If a drop of blood taken from an active well-fed Astacus is examined with a Zeiss Ob. D or E it is found to contain a large number of actively amoeboid corpuscles characterized by the possession of a great number of extremely large, highly refractive granules, or rather spherules (Fig. 1). If the preparation of the specimen has occupied a few seconds there will be seen, in addition to the spherule-bearing cells, a number of large, distinct, rounded nuclei which float free in the plasma. These latter elements belong to blood cells markedly distinct from the spherule-bearing corpuscles, and characterised by such an extreme sensitiveness to certain stimuli that contact with a foreign body, such as glass, causes an explosive disruption of their protoplasm. These cells, which I propose to call 'explosive corpuscles,' can be fixed by osmic vapour or iodine, and the study of such preparations (Fig. 3) makes very evident the fact that, as was pointed out by Heitzmann [note: '(9) ['Heitzmann. "Unters. ueber das Protoplasma." Sitzungsber. d. k. Acad. Wiss. in Wien, Bd. LXVII. 1873.']'] and Frommann [note: '(10) ['Frommann. "Unters. uber Struktur, Lebensercheinungen, und Reaktionen thieriseher u. pflanzlichen Zellen." Jen. Zeits. Bd. xvii. 1884.']] there are two distinct kinds of cells present in the blood of Astacus, both kinds. being amoeboid and both resembling the white rather than the red corpuscles of mammalian blood.' (166-167)

 

Figs. 2 and 8-9 in text:

'While the cell-substance is undergoing dissolution the nucleus also suffers remarkable changes. When the cell is first seen the nucleus is entirely invisible; but as the disintegration proceeds it rapidly comnes into view. The optical effect may best be likened to the rapid development of the image on a photographic plate. With almost the first stage in the dissolution of the cell protoplasm the nucleus becomes visible as a faintly-defined oval body. It rapidly acquires distinctness, the outline becomes firmer, and highly-refractive, yellowish-green masses appear, some of which join one another to form the periphery or "capsule" of the nucleus, while the others are distributed as rounded spherules or "nucleoli " in its interior (Fig. 2). At the same time traces of a network with large meshes may appear and rapidly vanish. This change which appears to be of the nature of a rigor or clotting, marking the death of the nucleus, converts the substance of the nucleus from an invisible matter, whose refractive index differs only slightly from its surroundings, into one of remarkable visibility, and is accompanied by an alteration in shape, the nucleus changing from an ellipsoid to a sphere, and expanding slightly (Figs. 8 and 9).' (169)

 

Figs. 3 and 11 in text:

'On studying the stained or unstained osmic preparation with the microscope we see that the morphological differences between the two types of cells are rather increased than diminished by the reagent. The explosive cell is not only the smaller btut its cell-substance is affected by the reagent in a way markedly different from that of the eosinophile cell. The cell-substance is clear, faintly visible and is marked by very fine granulations, or appears quite hyaline. In each cell is an oval nucleus containing an ill-defined and irregular network. The nuclear network is never perfectly preserved, the bulk of its substance has broken down to form nucleoli (Figs. 3 and 11). The most striking feature of these cells however is the great irregularity and diversity of form they exhibit.' (170-171)

 

Figs. 3 and 12 in text:

'Reaction with Osmic Vapour. A second and more valuable differential reagent is found in osmic vapour. As has been noted previously the eosinophile granules absolutely refuse to stain in the vapour, while the cell protoplasm becomes markedly distinct, so as to produce the appearance shown in Fig. 3.

If Fig. 3 is contrasted with the appearance of a basophile cell shown at Fig. 12, as it appears after treatment with osmic vapour, the profound difference of the two types of cells, a difference which exists in spite of their close similarity in the fresh condition, is at once obvious. The granules of the basophile cell after treatmient with osmic vapour stand out sharply distinct from the still clear hyaline cell-substance.' (178)

 

Fig. 3 in text:

'Methylene blue after osmic vapour stains both kinds of cells.

After treatment with osmic vapour the ectosarc of the eosinophile cells cannot be distinguished as a separate structure (Fig. 3) even though the preparation be deeply stained. The cells may however by fixation with a saturated solution of corrosive sublimate, be preserved with pseudopodia still extruded and the ectosarc clearly shewn as a hyaline layer surrounding the more prominent endosarc.' (175)

 

Fig. 5 in text:

'The endosarc... is more or less completely occupied by large granules or spherules. Sometimes it is partly occupied by granules and partly by vacuoles...

In some cases these cells assume a remarkable external resemblance to a Heliozoon such as Actinophrys, filiform and immobile pseudopodia radiating from the body of the cell. The resemblance is much heightened when the endosarc is relatively free from granules. The numerous vacuoles which may be present under those circumstances recalls with singular exactness the "bubbly" or vesicular natuire of the Heliozoon cell body (Fig. 5).' (172) 

'It may be noticed in passing that .25 % iodine prevents indefinitely the explosion of the corpuscles, the shell which remains after solution of the granules (Fig. 5) persisting unchanged. Also the iodine permanently fixes the granules and cell-substance of the eosinophile cells.' (181)

 

Fig. 6 in text:

'The blood corpuscles of Daphnia reproduce by direct fission. This fact has been previously noted by other workers. The blood corpuscles of Astacus also reproduce in a similar way. Fig. 6 shows an explosive corpuscle undergoing division.' (187)

 

Figs. 7-9 in text:

'The various stages in the disintegration or explosion of the explosive corpuscle may be fixed for prolonged examination by taking advantage of the preservative action of iodine. If a drop of the iodine solution be added to a drop of fresh blood the corpuscles are absolutely fixed, shewing the shape and for a time the granulations of life. If however a drop of iodine solution and a drop of blood are placed side by side on a slide and a coverslip lowered on to them so as to avoid blending them, we have, at the contact between the two drops under the coverslip, a narrow zone where they have mixed. In this zone the living and unchanged corpuscle has been fixed and preserved. Beyond this region and towards the blood the iodine has taken a longer and longer time to penetrate, and we thus get the corpuscles fixed at longer and longer intervals as we penetrate into the blood region. It was in this way that the stages shewn in Figs. 7, 8 and 9 were obtained.' (170)

 

Fig. 10 in text:

'Reaction with Sodium Sulphindigotate. This pigment serves as another instance of an acid pigment but of different chemical character. If it be dissolved in normal salt solution and added to blood the granules axe stained an intense blue (Fig. 10). The imbibition of the dye, when the living cells are exposed to its influence, takes place only at a critical point in the degradation of the molecular structure of the cell-substance. If the cell is possessed of great vitality it may be some hours before this point is reached.' (173-174)

 

Figs. 7 and 15 in text:

'Action of Iodine. This reagent furnishes us with a remarkable and stiggestive series of facts...  if we watch the cell after treatment with the reagent, we see a gradual disappearance of certain of its elements, namely, the large number of fine discrete granules. With 0.5 to 1% of iodine the cells resemble the specimen shewn at Fig. 7. The nucleus, as in the fresh cell when first seen, is invisible, being obscured by the large number of fine granules which are imbedded in the cell protoplasm and confer on it a characteristically solid, dense appearance. If only .25% of iodine be present these fine granules are at first present, but in from 10 to 15 minutes they are dissolved, the cell protoplasmn assuming the clear hyaline character so noticeable in preparations fixed by osmic vapour (Fig. 15). The impression conveyed by the brief glimpse one is able to have of the cell in the fresh state, that the protoplasm contains fine granules in considerable abundance, is confirmed by the appearances presented after treatment with iodine.' (171)

 

Fig. 12 in text:

'The basophile cells are not only characterised by the reactions of their spherules and cell-substance with basic pigment and osmic vapour; they also present certain definite and peculiar structural features. They are of irregular shape, being either square, triangular, or rounded, and in size are larger than the eosinophile cells. The basophile cell shown at Fig. 12, and detected in blood drawn from the pericardial sinus is squarish in outline. Each side of the square measured 38µ and the nucleus was 120µ in diameter. I have never detected any trace of amoeboid movement in a basophile cell, though it is probable that there are sluggish movements, which from being very slow and slight and occurring at several points of the surface at the same time, escape observation.' (178)

 

Fig. 13 in text:

'The amorphous masses are apparently derived from the granules by their fusion. This is well shown in Fig. 13, where the outlines of the granules are still imperfectly retained. This fusion of the granules is part of the normal process going on in the cell, the process however is not a simple blending of the granules but their substance at the same time largely loses its basophile reaction, staining only very slowly and with difficulty. When the staining does take place however the tint is pronounced and of a rose colour with yellow light.' (179)

 

Fig. 14 in text:

'Leaving the connective-tissue framework, which is of no immediate interest, we pass to the second constituent of the tissue, the enclosed cells.

The cells are large and irregular in shape. They vary in size, the one shown at Fig. 14 was found to have the extreme dimensions of 65µ in length and 37µ in breadth. In their general character and reactions these cells accurately resemble the basophile cells occasionally found free in the blood. Frequently however they contain large spheres, which by their reaction with osmic acid and froin the fact that they are soluble in such reagents as turpentine we may conclude to be fat.

If this tissue, immediately after its removal from the animal, be slightly teased in a very dilute solution of methylene blue and then examined in the course of half-an-hour to an hour, with strong yellow light the cell granules will appear as shown in Fig. 14, that is, they show a marked rose colour, at the same time the nuclei of the cells and the nuclei of the connective tissue appear blue, like the nuclei of the basophile cells when stained. In other words we see that the spherules of this tissue, like the spherules of such basophile cells as may be found free in the general circulation, give this peculiar "rose reaction" with inethylene blue and yellow light.' (180)

 

Fig. 16:

'I used for the purpose of the experiments Indian ink, and it was injected suspended in sterilised (boiled) normal salt solution [note: 'Control experiments were carried out to determine the effect of normal salt solution alone when injected in varying amounts. They do not affect what is stated above and therefore will be passed by for the present.']... 

If the blood be examined in the fresh condition about an hour after the injection the plasma will be found to be still highly charged with ink particles. Here and there in the preparation large cells are to be seen (Fig. 16) of an irregular shape, with numerous irregular and exceedingly fine pseudopodia, and with striking vacuoles of varying size in their substance, which is of a very clear transparent nature. These cells are highly charged with ink particles which are imbedded in patches in the cell-substance, in the neighbourhood of a vacuole.' (182-183)

 

Fig. 17-18 in text:

'What is the origin of the above-mentioned large hyaline cells? The study of osmic vapour preparations leads to the conclusion that they are nmodified explosive cells, formed in some cases at any rate by the fusion of two or more to form a plasmodium. In the osmic vapour preparation we notice that the great number of the explosive cells are charged with ink particles (Fig. 18). Where this is not the case we see a striking incipient vacuolation (Fig. 17). On the other hand the eosinophile cells show no included particles.

We thus have the remarkable fact that power of ingesting solid particles of the nature of Indian ink appears to reside only in one of the two kinds of cells normally found in the blood of Astacus. On the other hand, and the fact has a certain bearing on the function of the basophile cells, the cells of the basophile "Zellgewebe" take in the particles of indian ink to a limited extent. Every cell in the tissue may, and probably does, contain ink particles but never in great abundance. In the place of dense clumps of aggregated granules we find scattered, isolated ink particles which are grouped round the basophile granules.' (183-184)

 

Fig. 24 in text:

'Daphnia possesses a special fat-holding tissue which is composed essentially of rounded cells anchored by fine processes. Under the circumstances just mentioned these become charged with fat particles. I further convinced myself that this tissue may, when a great quantity of fat is absorbed, be recruited from the blood cells which fix themselves as now stationary fat-holding cells. In Fig. 24 a recently fixed corpuscle is shown.' (187)