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Date: 1894
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Cited by A.A. Kanthack and W.B. Hardy, 'The Morphology and Distribution of the Wandering Cells of Mammalia', Journal of Physiology 17 (1-2) (1894), pp. 80-119.
Description:Explanation of Plate II (figs. 1-28):
All the figures are to the same scale and drawn with Oc. 4, Obj. 1th how. imm. with a camera lucida.
Figures 1, 2, 3, 4 and 5 Human.
Fig. 1. Cells of blood of healthy boy. Fluid preparation made with the methylene blue solution. (a) coarsely granular oxyphile cell, (b) finely granular oxyphile cell, (c) hyaline cell, (d) lymphocyte, (e) and (f) finely granular basophile cell.
Fig. 2. Blood from same boy. Film preparation exposed to a cold saturated solution of eosine in 50 per cent. spirit for about 5 seconds, then to Loeffler's methylene blue. (a) coarsely granular oxyphile cell, (b) finely granular oxyphile cell, (c) hyaline cell. Fig. 3. Blood from healthy adult. Film preparation stained with the " neutral " mixture described on page 84. (a) coarsely granular oxyphile cell, (b) finely granular oxyphile cell, (c) hyaline cell.
Fig. 4. Coarsely granular basophile cell, connective tissue. Human.
Fig. 5. Lymphocyte to show nuclear network. Flemming's fluid. Haematoxyline.
Fig. 6. Rat. Cells from blood. Film preparatioin stained with glycerine eosine and Loeffler's methylene blue. (a) coarsely granular oxyphile cell, (b) finely granular oxyphile cell (note the complete absence of staining in the granules, cp. with Fig. 11), (c) hyaline cell.
Fig. 7. Rat. Cells of peritoneal fluid. Film preparation stained with glycerine eosine and Loeffler's methylene blue. (a) adult and (a') young coarsely granular oxyphile cell, (b) adult and (b') young coarsely granular basophile cell, (c) adult and (c') young hyaline cell.
Fig. 8. Rat. Cells of peritoneal fluid. Fluid preparation made with the methylene blue solution. (a) coarsely granular oxyphile cell, (b) hyaline cell, (c) young form of coarsely granular basophile cell. The adult form of the coarsely granular basophile cell is omitted and -
Fig. 9. The coarsely granular basophile cell of the mouse is given in its place. From a fluid preparation made with the methylene blue solution.
Fig. 10. Rat. Cells from a film of subcutaneous connective tissue, stained with glycerine eosine and Loeffler's methylene blue. (a) coarsely granular oxyphile cell, (b) coarsely granular basopliile cell.
Fig. 11. Rat. Finely granular oxyphile cell from film of blood stained with the "neutral " mixture as described on page 84.
Fig. 12. Guinea-pig. (a) and (b) coarsely granular oxyphile cell, and hyaline cell from a fluid preparation of peritoneal fluid made with the methylene blue solution. (c) finely granular oxyphile cell from fluid preparation of blood made with the methylene blue solution. To show the differences between the oxyphile cell of the peritoneal fluid and the finely granular oxyphile cell of the blood. (d) coarsely granular oxyphile cell from a film of peritoneal fluid stained with an aqueous solution of sodium sulphindigotate. A very much flattened cell was chosen for drawing in order to show clearly the individual granules and the clear unstained space which represents the nucleus.
Fig. 13. Guinea-pig. Coarsely granular basophile cell in various stages of disintegration. Film preparations stained with glycerine eosine and Loeffler's methylene blue. (a) from wall of peritoneal cavity, fixed with absolute alcohol. (b) from film of peritoneal fluid, fixed by heat. (c) and (d) from film of subcutaneous tissue, fixed by drying at temperature of room.
Fig. 14. Rabbit. Coarsely granular oxyphile cell from film of peritoneal fluid stained with glycerine eosine and Loeffler's methylene blue.
Fig. 15. Rabbit. Cells of blood shortly after a heavy meal. Fluid preparation made with the methylene blue solution. (a) coarsely granular oxyphile cell, (b) finely granular oxyphile cell, (c) hyaline cell, (d) lymphocyte, (e) finely granular basophile cell.
Fig. 16. Rabbit. Blood; fluid preparation made with the methylene blue solution. About 10 hours after a meal. (a) finely granular basophile cell, (b) lymphocyte.
Fig. 17. Rabbit. Coarsely granular basophile cell from subcutaneous connective tissue stained with the methylene blue solution.
Fig. 18. Rabbit. Finely granular basophile cell, inflamed area stained with the methylene blue solution.
Fig. 19. Film of subcutaneous connective tissue taken from the neighbourhood of a Ziegler's chamber filled with diluted broth culture of the comma bacillus seven hours after introduction of the chamber. Guinea-pig. Eosine in 90 per cent. alcohol, Loeffler's methylene blue, Oc. 4, Ob. D, cam. luc. A part of the preparation where the coarsely granular oxyphiles were relatively few in number was chosen for the drawing in order to show the connective tissue elements.
Fig. 20. Film of subcutaneous connective tissue from normal guinea-pig for comparison with fig. 19.
Fig. 21. Living cells attacking a chain of Bacillus ramosus. Ziegler's chamber 2 1/2 hours in peritoneal cavity of guinea-pig. The chamber was removed unopened to warm stage and examined with Oc. 4, Ob. D.
Fig. 22. Hanging drop of blister fluid from forearm inoculated with Bacillus ramosus. (a) coarsely granular oxyphile cell at rest, hanging drop at temp. of room. Warm water was then allowed to run into warm stage, cell became active and applied itself to bacilli - (b) and (c). Human.
Fig. 23. Blister fluid inoculated with Bacillus ramosus. (a) chain of bacilli with three cells applied to it, Oc. 2, Ob. A, (b) part of the same chain stained with the methylene blue solution, Oc. 4, Ob. 1/12th, cam. luc. Human.
Fig. 24. Hanging drop of blister fluid. Phagocyte with ingested fragments of Bac. anthracis, Oc. 4, Ob. D, cam. luc. Human.
Fig. 25. Peritoneal fluid, rat. 15 minutes after injection of Bac. pyocyaneus. Plasmodial mass of coarsely granular oxyphile cells with surrounding hyaline cells. Oc. 4, Ob. 1/12th, cam. luc.
Fig. 26. Same experiment as last. Single cell attacking bacilli. Oc. 4, Ob. 1/12th, cam. luc.
Fig. 27. Peritoneal fluid, rat. 10 minutes after injection of bac. anthracis. Fluid preparation stained with the methylene blue solution. Part of much distorted anthrax chain attacked by oxyphile cell and also being ingested by phagocyte.
Fig. 28. Peritoneal fluid, rat. 15 minutes after injection of anthrax. Coarsely granular oxyphile cell which has discharged its granules. Fixed by heat. Oc. 10, Ob. 1/12th apochr. Powell and Lealand.' (108-110)
Figs. 1-3, 6-8, 12 and 15 in text:
'The Hyaline Cell. Figs. 1, 2, 3, 6, 7 c, 8 and 12 b, and 15 c. This is usually of about the same size as the coarsely granular oxyphile cell and rather larger than the finely granular cell. When at rest it is a spherical cell but, owing to its pseudopodial activity, it mostly appears as a somewhat tenuous body of very irregular shape. The nucleus is usually spherical, it is sometimes kidney-shaped probably as a result of the mode of killing the cell. A very fine nuclear network with large meshes spreads through the nucleus and, in most preparations, a nucleolus is present. The cell substance is always free from discrete granules, it is not, however, very transparent but presents rather the appearance of ground glass.' (96)
Figs. 1-2, 6-8, 10, 12 and 14 in text:
'The coarsely granular oxyphile cell, or eosinophitle cell (Figs. 1, 2, 6; 7, 8, 10, 12 (a), and 14), varies in size in different animals, not only absolutely, but relatively to the dimensions of the other classes of cells. In man it is larger than either the hyaline cell, the finely granular oxyphile cell, or the finely granular basophile cell. In the rat, rabbit, and guinea-pig, on the other hand, it is smaller than the largest hyaline cells, but larger than the finely granuilar oxyphile and basophile cells [note: 'The dimensions of the various cells are given in the table on p. 101.']. (88-89)
Figs. 1-2, 6, 11-12 and 15 in text:
'The finely granular Oxyphile cell (Figs. 1, 2, 3, and 6 b, 11, 12 c, and 15 b), is always smaller than the coarsely granular oxyphile cell. On the other hand, it is in man and the guinea-pig rather smaller than the hyaline cell, and in the rat and rabbit very considerably smaller. The nucleus is an exceedingly irregular structure branching throughout the cell (Figs. 2 and 6 b). The branches swell out here and there into masses of an irregular shape, the intervening or connecting portions being of the nature of slender bars or threads. A fine and close nuclear network can be detected.' (89-90)
Figs. 1, 5, 7-8 and 15 in text:
'Three forms of immature cells are readily recognised in the various body fluids of a normal animal. These are as follows:-
(i) A small round cell characterised by possessing a deeply staining horse-shoe shaped nucleus embedded in scanty cell substance. The cell substance is charged with minute granules which give the same reactions as the coarse oxyphile granules (Fig. 7a'), that is to say, using the ordinary terminology, they are undoubtedly eosinophile granules...
(ii) A round cell, somewhat larger than the last (diameter 8 to 9 µ). It possesses a spherical nucleus and the cell substance is charged with small basophile granules (Figs. 7b', 8c). It is readily distinguished from the finely granular basophile cell of the blood by its nucleus, which is a simple sphere instead of being trilobed, and by its granules, which are larger, each being a distinct though small spherule...
(iii) A small cell which, like the first described, is 6.5 µ, in diameter (rat) and possesses a deeply staining nucleus and scanty cell substance (Figs. 1d, 15, 7c', 15d). The nucleus, however, is spherical and the cell substance is free from granules. These cells have been called lymphocytes, from the fact that they are produced in lymphatic glands.' (98)
Figs. 1, 6, 12 and 15 in text:
'The finely granular basophile cell (Figs. 1 e and f, 6 d, 12 e, 15 e) unlike the last, is a small cell, being usually the smallest of all the wandering cells. It is spherical in shape and possesses a characteristically trilobed nucleus. The cell-substance is clear and optically structureless, and usually contains an immense number of mirnute granules which often appear as mere points, and are characterised by staining a very opaque blue or purple colour with methylene blue. Another very characteristic feature is the fact that the cell substance colours a purple or pink tinge with the methylene blue solution.' (94)
Figs. 1, 8 and 12 in text:
'Cell granules. The cell granules are relatively large, spherical, or slightly ovoid bodies, and are sharply marked off from the cell substance by their very high refractive index, which is so great that in fluid preparations the granules have a brilliant greenish lustre (Figs. 1, 8, and 12 a). The cell-substance in which they are imbedded has the appearance of a clear transparent structureless jelly.' (89)
Figs. 1 and 12 in text:
'The granules are very small spherical bodies which crowd the otherwise clear and optically structureless cell-substance. They have a refractive index only slightly above that of the medium in which they are imbedded so that they are scarcely visible when unstained (Figs. 1 b and 12 c). Their oxyphile reaction is much feebler than that shewn by the large granules of the coarsely granular oxyphile cells.
Figs. 4, 7, 9-10 and 13 in text:
'The coarsely granular Basophile cells (Figs. 4, 7, 9, 10, 13, and 17) have been described by Ehrlich [note: 'Ehrlich. Loc. cit.'] under the name of 'Mastzellen' and their existence in the coelomic spaces and in the connective tissues was recognised by him and by Ranvier [note: 'Ranvier. Comptes Rendus, T. cx. p. 768.']. The cells of the connective tissues and those of the coelomic fluid differ slightly in size and shape, and we will therefore deal with them separately.' (92-93)
Figs. 7-8, 12 and 14 in text:
'The coelomic fluid of mammals is very richly supplied with wandering cells, indeed it presents from this cause a cloudy turbid appearance. Of the cells present from 30 to 50 per cent. are coarselv granular oxyphile cells (Figs. 7, 8, and 12 a and 14). In connective tissue again, the total number of these cells must be very great, for the thinnest film spread out on a slide will frequently show from 5 to 10 in a single field of a Zeiss D, Oc. 4. Thus this cell has an extraordinarily wide distribution outside the vascular system.' (91)
Figs. 7 and 9 in text:
'The coarsely granular basophile cell found in the coelomic fluid is, so far as we know, under normal conditions not endowed with the power of amoeboid movement. It is a large cell having the form of a very much flattened sphere which might not inaptly be compared with a millstone whose edges had been very much rounded off (Figs. 7 b, and 9). The nucleus is rounded, occupies a central position and seems to be singularly devoid of chromatin, staining with difficulty. The cell substance, clear and optically structureless, is charged with a very great number of exceedingly large spherules, the basophile "granules."' (93)
Figs. 7 and 10 in teext:
'The nucleus is typically an elongated body bent to form a horseshoe. In the rat the arms of the horse-shoe are carried so far round that in film preparations the ends often overlap, giving to the nucleus the appearance of a circle with a large hole in the centre (Figs. 7 and 10 a). Sometimes the nucleus is lobed, but we are inclined to regard this appearance as being largely due to the stresses to which the nucleuis is subjected when the cell is dying. In the living cell at rest, when it is spherical, the shape of the nucleus, so far as it can be determined by the disposition of the cell-granules, is a simple horse-shoe or crescent.' (89)
Figs. 8 and 12 in text:
'The cell substance of the hyaline cells differs, at first sight, from that of the granule bearing cells, in taking a faint diffuse colouration with basic dyes. The highest powers of the microscope, however, frequently resolve this cloudy and apparently continuous colour into a number of stained points (Figs. 8 and 12 b); and the cell substance of the dead hyaline cell is then seen to be composed of an optically structureless substance remarkably indifferent and resistant to all stains, embedding a staining substance dispersed throughout it in the form of a cloud of minute particles.' (96)
Figs. 10, 13 and 17 in text:
'The coarsely granular basophile cell of the connective tissue (Figs. 10 b, 13 a, c, d, and 17) differs from the other forms of wandering cells in that it appears to be, at any rate in the normal animal, not only nonamoeboid like its coelomic brother but also stationary. It is therefore, except perhaps under certain conditions, not strictly a wandering cell. It is of a rounded or slightly polygonal shape and usually is more flattened, and so rather larger in its extreme dimensions, than the cell of the coelomic fluid. In its other histological features, such as the nature of its granules, nucleus, and cell substance, it exactly resembles the coelomic cells.' (93)
Fig. 12 in text:
'In preparations of the blood of the guiinea-pig made with the methylene blue soltution, cells are found like the one shown in Fig. 12 e. The granules are large and stain with the metbylene blue, they are therefore, at least under certain conditions, basophile. It is this cell which appears in Table II. as the finely granular basophile cell of the guinea-pig. We are not, however, at all certain that it is really identical with the very characteristic and readily detected cell of man and the rabbit. Our knowledge of the finely granular basophile cell, however, is so deficient that it is perhaps wisest for the present to class it as we have done.' (95)
Fig. 13 in text:
'Cells characterised by great instability have been described elsewhere in Astacus [note: 'Hardy. This Journal, xiii. Nos. 1 and 2.'] as the "explosive" cell, of that animal, and the basophile cells of the guinea-pig and rabbit might, with equal justice, be designated the explosive cells of those animals. Under the influence of certain chemical stimuli, as will be seen later, the basophile cells of the rat also become explosive. The instability of the coarsely granular cells of the guinea-pig and rabbit is so marked and constant a feature that Ranvier was unable to detect their presence in the coelomic fluid of these animals, and came to the conclusion that they were absent [note: 'Ranvier. Comptes Rendus, cxix., footnote p. 923.']. Examples from the guinea-pig are shown in Fig. 13, (a) being from the wall and (b) from the fluid of the peritoneal cavity, while (d) and (c). represent the appearance produced by the bursting of the cells in a film of connective tissue.' (94)
Figs. 19-20 in text:
'If we ask ourselves the question where do these cells come from we are able to answer positively, in the case of chambers placed under the skin, that they have their immediate origin in the surrounding connective tissue. Fig. 19 shows the striking appearance presented by a film of connective tissue taken from the immediate neighbourhood of a Ziegler's chamber filled with a dilute broth culture of the cholera vibrio after 7 1/2 hours' sojourn under the ventral skin of a guinea-pig. The chamber was found to contain multitudes of the coarsely granular oxyphile cells, and a comparison of figure 19 with figure 20 will show how strikingly abundant were these cells in this leucocytic focus as compared with the normal connective tissue.' (104)
Fig. 20 in text:
'The most striking change was found in the connective tissue which formed the walls of the space in which the chambers were placed, for this was packed with an immense number of the coarsely-granular oxyphile cells together with a smaller number of the coarsely-granular basophile cells. A teased-out film of areolar connective tissue (Fig. 20) taken from a normal animal is seen to consist, so far as bulk is concerned, mainly of the fibrous matrix.' (107)
Fig. 21 in text:
'Tubes or Chambers in the Peritoneal Cavity. Fine capillary tubes were used and were filled with very dilute nitrate of silver, or turpentine, or with the diluted filtrate from a broth culture of pyocyanine which had been passed through a Chamberland's filter...
The chambers were inserted through a slit about 3/4 inch long made in the linea alba; the wound was then sewn up and dressings applied. They were allowed to remain in the peritoneal cavity for 2, 2 1/2 or 7 hours. The animal was then killed and the chambers were at once removed and the cells on the external surface and those in the interior were examined at once and without the addition of any reagent. In this way the cells were seen to perform amoeboid movements, and to be attacking the bacilli (Fig. 21). The chambers were then split open and the cells adherent to the external surface and those in the interior were examined with the aid of the methylene blue solution, and as film preparations.' (104-105)
Fig. 24 in text:
'The hyaline cells unlike the coarsely-granular oxyphile cells manifest a true phagocytosis, i.e. the intussusception of discrete particles into their substance, and their solution in digestive vacuoles (Fig. 24 human).' (112)
Figs. 25 and 27 in text:
'In a former paper [note: 'Kanthack and Hardy. Loc. cit.'] we described the formation of plasmodial masses of wandering cells in the course of the conflict with the micro-organisms. Such bodies are readily seen in hanging drops of blister fluid, in Ziegler's chambers, or in preparations of peritoneal fluid containing bacilli. Fig. 27 shows a striking case, a chain of anthrax being attacked by a coarsely granular oxyphile cell at one place, while at the same time it is being ingested by a phagocyte at another point. Fig. 25 shows an interesting stage, the oxyphile cells being massed in the centre of a number of hyaline cells.' (113)