222 Hematologic Elements and Related Issues (chap. 16) 



Solution B: 



eosin Y, C.I. 45380 I.O gm. 



dibasic sodium phosphate, anhydrous 5.0 gm. 



monobasic potassium phosphate 6.25 gm. 



distilled water 500.0 ml. 



Dissolve phosphate salts first, then add stains. (The azure will go into 

 solution by grinding in glass mortar with a small amount of phos- 

 phate solvent.) Set aside for 24 hours. Filter. 



procedure: 



1. Dip slides in solution A: 1 second. 



2. Rinse gently in clean distilled water: few seconds, or until stain 

 ceases to flow from film. 



3. Dip in solution B: 1 second. Use this solution with care; it tends 

 to decolorize the leukocytes stained with methylene blue-azure, 

 and accelerates the dissolving of hemaglobin. 



4. Rinse gently in distilled water: 2-3 dips. 



5. Dry, do not blot. Stand slides on end and allow to air dry. 



6. When completely dry, add mounting medium and cover glass. 



results: 



There will be a thicker central area of partially laked blood. This 

 may not be well-suited for examination. But the surrounding area 

 and especially at the edge toward which the hemaglobin drained will 

 be creamy, sometimes mottled with pale blue. This is the best area 

 for study. 



leukocytes: 



cytoplasm — pale blue, poorly defined 



nuclei — dark blue, well defined 



eosinophilic granules — bright red, large, well defined 



neutrophilic granules — pale purple pink, small indistinct 



basophilic granules — deep blue with reddish cast 



platelets — pale purple or lavender 



parasites: 



cytoplasm — blue 



chromatin — purplish red or deep ruby red 



pigment — unstained yellow granules of varying intensity 



