1908.] 



Diffusion into Living Cells. 



103 



network stained better if its concentration was increased. In order to obtain 

 staining of the nuclei of lymphocytes in 10 minutes on agar from a tube 

 which contained 4 units, instead of 2 units, of stain, the amount of alkali 

 added was of course less in proportion to the increased concentration. With 

 4 units of stain, the other factors, except alkali, being as before, the equation 

 now stood as : — 



c/= (4s + Qa+%+t)- (3c + n) = 14. 



The red corpuscles appear to have a very high cf. I caused them to stain 

 on agar which contained 1 c.c. of stain (10 units) and 11 units of alkali, in 

 the presence of 1-per-cent. sodium citrate and 0'8-per-cent. sodium chloride 

 at 37° C. in 10 minutes. This was the equation : — 



cf= (10s+lla + 7h+t)-(3c+n) = 25* 



jExamples. — A growth of staphylococci had a cf of 16. 



How much alkali must 10 c.c. of jelly contain to cause the germs to 

 stain in 10 minutes if the jelly already contains 5 per cent, of stain, 15 per 

 cent, sodium citrate, and 0'8 per cent, sodium chloride, when the slide is 

 incubated at 37° C. ? 



a = (16c/+4-5c + ?i)— (5s+7A + 0, 



a = 8 - 5 units, or - 0425 gramme of sodium bicarbonate. 



A strain of typhoid bacilli had a cf of 21. A tube of agar contained 6 units 

 of alkali solution and the usual quantities of sodium citrate and chloride. 

 How much stain should be added to the tube to produce staining of the 

 bacilli in 20 minutes at 37° C. ? 



s= (21cf+3c + n)-(Qa+7h+2t), 



s = 10 units of stain, i.e., 1 c.c. 



* I do not think this is strictly accurate, for it depends on the coloration of the 

 stroma. Nucleated red cells have a comparatively low cf, resembling that of the poly- 

 morphonuclear cells, though I am also doubtful of this point, because I have only been 

 able to obtain these nucleated cells from persons suffering from disease, and, as I have 

 already shown (8), all the blood cells in chronic illnesses, especially phthisis, malaria, and 

 Hodgkin's disease, have so far shown a general fall in their coefficient of diffusion. 

 Again, attention may be drawn to the comparatively low cf of the granular erythrocytes 

 which constitute about 1 per cent, of all the red cells in healthy blood as demonstrated 

 by this method of examination (1). The rate of staining of the granules is about the 

 same as those of the polymorphonuclear leucocytes, but the rate of staining of the stroma 

 of these granular cells is much higher than that of their granules, yet lower than the rate 

 of staining of the stroma of ordinary erythocytes. This fact is the more interesting, 

 because it is almost in these granular red cells alone, which have a low cf, that " red 

 spots " are to be seen, although I have on three occasions seen them in ordinary red cells, 

 and the spots appear to be dependent on diffusion (4). These cells also become achromatic 

 more readily than ordinary erythrocytes. The granules have been described (1) as the 

 remains of a nucleus. The question, therefore, of the cf of the red cells I will leave for 

 the present as indicated. 



