1770 Journal of Applied Microscopy 



By this method the red cells are pale pink or greenish in tint and semi-trans- 

 parent ; the polynuclear leucocytes show a nuclear network stained a ruby red 

 color with sharply defined margins, and a colorless, extra-nuclear protoplasm 

 with fine red granules ; the mononuclear leucocytes have ruby red nuclei with 

 extremely sharp, clear margins and a pale eau de-Nil or blue extra-nuclear proto- 

 plasm with occasional red granules ; the lymphocytes are the same as the mono- 

 nuclears except their nuclei are generally more deeply stained ; the eosinophilic 

 leucocytes exhibit a ruby red nucleus and pale pink granulations. The granules 

 of the basophiles are very densely stained a deep purplish black ; the nuclei are 

 more or less masked by the granules which overlie them. The nucleated red cells 

 show a nucleus almost black, with sharp outlines and a grey extra-cellular por- 

 tion. The blood plates are of a deep ruby red with a pink margin. They fre- 

 quently exhibit a pale blue peripheral zone surrounding the red center. The 

 bodies of the malarial parasites are stained blue while the chromatin particles 

 take on a ruby red color. 



Wright (The Journal of Medical Research, January, 190'2) has modified the 

 above method. He thus prepares his staining fluid: A one-half per cent, watery 

 solution of sodium carbonate is made in an Ehrlenmeyer flask and to it is added 

 a one per cent, methylen blue (Griibler) solution. This mixture is to be steamed 

 in an Arnold sterilizer for one hour. It is then cooled and without filtering is 

 poured into a large disk or flask and a sufficient quantity of a 1 in 1(100 solution 

 of eosin (Griibler, yellowish, soluble in water) is added till the mixture loses its 

 blue color and a scum with yellowish metallic lustre forms on the surface, while 

 on close inspection a finely granular black precipitate appears in suspension. 

 (This requires about 500 c. c. of the eosin solution for 100 c. c. of the alkaline 

 methylen blue solution.) The precipitate is collected on a filter and allowed 

 to dry without washing. When thoroughly dry a saturated solution is made of 

 this powder in pure methyl alcohol. (Three-tenths of a gramme of this precipi- 

 tate will thoroughly saturate 100 c. c. of methyl alcohol in a few minutes.) This 

 saturated alcoholic solution is next filtered and twenty-five per cent, of methyl 

 alcohol is added to the filtrate. The resulting fluid is the staining fluid, which 

 keeps well. Care, however, should be taken that the alcohol should not 

 evaporate. 



Wright's staining method is somewhat different from Leishman's. After the 

 blood films are made in the usual way, as much of the stain is added to the 

 preparation as will not drain off. This is allowed to remain one minute on the 

 slide or cover-glass before dilution. By this process the blood films are fixed. 

 Water is then added drop by drop till the mixture becomes semi-translucent and 

 a yellowish scum forms on the surface. This mixture should remain on the 

 preparation for two or three minutes. By it the real staining takes place. At 

 the end of this time the preparation is washed in water till the film assumes a 

 yellowish or pinkish color in its better spread portions. Care should be taken 

 lest the decolorization in water goes too far. As a rule this process takes one 

 to two minutes. The specimen is now dried between filter paper and mounted 

 in balsam. 



The color reactions obtained by this method differ from Leishman's. Instead 

 of a ruby red nucleus a dark lilac or blue one is seen ; the granules of the 

 polynuclear leucocytes are of a reddish lilac color. The red blood cells are 

 orange or pink in color. For a more detailed account of the microscopical ap- 

 pearances of blood-films stained by this method one is referred to the original 

 article. w. r. s. 



