228 THE CIRCULATORY SYSTEM. 



stain (preferably eosin) in glycerin, washed with water, subsequently col- 

 ored with a nuclear stain (as hernatoxylin or methylene-blue), and then 

 dried and mounted in Canada balsam. Sections may be treated in the 

 same way, with the exception that after being washed with water, they 

 are first dehydrated with absolute alcohol before mounting in balsam. 



Another method by which both nuclei and granules are stained 

 consists in the use of Ehrlich's hematoxylin solution (see page 43), 

 to which 0.5% eosin is added. Before using, the solution should 

 be permitted to stand exposed to the light for three weeks. This mixture 

 stains in a few hours, after which the preparation is washed with water, 

 treated with alcohol, and then mounted in Canada balsam. The 

 a -granules appear red, the nuclei blue. 



The /^-granules (amphophile, indulinophile) stain as well in 

 acid as in basic anilins. They do not occur in man, but may be observed 

 in the blood of guinea-pigs, fowl, rabbits, etc. They are demonstrated 

 as follows : Equal parts of saturated glycerin solutions of eosin, naph- 

 thylamin-yellow, and indulin are mixed, and the dried preparations treated 

 with this combination for a few hours, then washed with water, dried 

 between filter-paper, and mounted in Canada balsam. The amphophile 

 granules are stained black, the eosinophile granules red, the nuclei black, 

 and the hemoglobin yellow. 



The ^-granules, or those of the mast-cells, are found in normal 

 tissues and also in small quantities in normal blood, and are found in 

 larger numbers in leukemic blood. They may be shown by two methods : 

 ( i ) A mixture is made consisting of concentrated solution of dahlia in 

 glacial acetic acid 12.5 c.c., absolute alcohol 50 c.c., distilled water 100 

 c.c. (Ehrlich). The treatment is the same as for the amphophile gran- 

 ules ; (2) Westphal's alum-carmin -dahlia solution (vid. Ehrlich). This 

 mixture is used in staining dry preparations as well as sections of objects 

 fixed for at least one week in alcohol. Alum i gm. is dissolved in dis- 

 tilled water 100 c.c., and carmin i gm. added. The whole is then 

 boiled for one-quarter hour, cooled, filtered, and 0.5 c.c. of carbolic 

 acid added (Grenacher's alum-carmin, see page 42). This solution is 

 now mixed with 100 c.c. of a saturated solution of dahlia in absolute 

 alcohol, glycerin 50 c.c., and glacial acetic acid 10 c.c., the whole stirred 

 and allowed to stand for a time. The specimen is stained for twenty-four 

 hours, decolorized in absolute alcohol for the same length of time, and 

 finally mounted in Canada balsam. The ^-granules are colored a dark 

 blue and the nuclei red. A simpler method of demonstrating the 

 ^-granules consists in overstaining dry and fixed cover-glass preparations 

 with a saturated aqueous solution of methylene-blue, decolorizing for some 

 time in absolute alcohol, drying between filter-papers, and mounting in 

 Canada balsam. 



The ^-granules (basophile) occur in mononuclear leucocytes 

 of the human blood. Their staining may be accomplished in a few min- 

 utes by treating fixed cover-glass preparations with a concentrated aqueous 

 solution of methylene-blue, after which they are washed with water, dried 

 between filter-papers, and mounted in Canada balsam. 



The e- or neutrophile granules which are found normally in 

 the polynuclear leucocytes of man (as also in pus-cells), in some of the 

 transitional cells, and in the myelocytes, are stained by Ehrlich as follows : 

 5 vols. of a saturated aqueous solution of acid fuchsin are mixed with i vol. 



