60 The Philippine Journal of Science isis 



with heat, in pure methyl alcohol or in equal parts of absolute 

 alcohol and ether. The slides were stained with eosin and 

 methylene blue, Ehrlich's triacid mixture, Jenner's stain, and 

 Wright's stain. Jenner's stain was used in making the differen- 

 tial counts, as it is easily handled, and gives very accurate 

 results. The size of the corpuscles was obtained with a Zeiss 

 ocular screw micrometer, using stained films, and taking those 

 parts of the film in which the cells were not crowded. 



The technique used in staining the preparation was as follows : 



When Jenner's stain was used, the filmi was dried in air and 

 flooded with the dye, which was left to act for from two to three 

 minutes. It was then washed from ten to fifteen seconds in 

 distilled water, and dried as rapidly as possible in air. When 

 well stained and washed, the red cells had a terra-cotta color. 



With Wright's stain, the film was dried in air, and flooded 

 with the dye which was allowed to act for one minute. Distilled 

 water was then added, drop by drop, until a film of metallic 

 luster began to form on the surface. This was allowed to act 

 from two to three minutes longer. The preparation was then 

 washed for ten seconds in distilled water, and dried in air as 

 rapidly as possible. With this stain, the red corpuscles took a 

 pinkish color. 



With eosin and methylene blue, the film was previously fixed, 

 either with heat or by submerging in pure methyl alcohol for ten 

 minutes, or in equal parts of absolute alcohol and ether for 

 ten minutes. Then it was flooded with saturated alcoholic solu- 

 tion of Ehrlich's blood eosin for about ten seconds and washed 

 in water. After this it was flooded with a saturated aqueous 

 solution of Grubler's methylene blue for one minute, washed 

 quickly with distilled water, and dried in the air. 



With Ehrlich's triacid mixture, the film was previously fixed 

 by placing the slides, film side down, upon a heated copper tray, 

 and held at a temperature just below boiling for fifteen minutes. 

 The film was then covered with the mixture and allowed to act 

 ten minutes, after which it was washed hastily in distilled 

 water and dried rapidly in air. 



The red corpuscles in the fresh condition appear as biconcave 

 disks of a homogeneous appearance, yellowish in color, and with 

 a nearly translucent central area and have the general ap- 

 pearance of human blood. With eosin and methylene blue and 

 with Wright's stain, the red corpuscles take a pinkish stain, 

 with Jenner's they are terra-cotta color, and with Ehrlich's 

 triacid mixture they have an orange tint. 



Five varieties of leucocytes were noticed in the circulating 



