Sex Chromatin 363 



2. Dilute with Wright's buffer (equal to amount oi stain); blood 

 smears: 30 seconds; oral mucosal and vaginal smears: 45 seconds. 



3. Wash in running water: 2-3 minutes. 



4. Decolorize in 50%, 70%, 95%,, and absolute alcohol: 30 seconds 

 each. 



5. (a) Air-dry blood smears and they are ready for use. 



(h) Clear oral mucosal and vaginal smears in xylene, 2 changes, 

 and mount. 



results: 



Staining: chromatin — blue 



Counting: oral mucosal and vaginal smears — 6 cells out of 100 

 coimted must be positive for female sex chromatin to be consid- 

 ered female. If less than 4 per 100, the specimen is male, 

 blood smears — at least 6 rich-staining "drumsticks" must be 

 counted in 500 polymorph cells for a female count. No more than 

 2 drumsticks should appear in a male count. The tiny clubs and 

 sessile nodules are found in about equal numbers in both sexes, 

 but cannot be counted as sex chromatin bodies. (See excellent pic- 

 tures in Greenblatt and Manaotou) 



comments: 



For sex chromatin counts, the preparation of a "huffy coat" is 

 recommended by the present author. It gives the examiner a fighting 

 chance at a relatively easy cell count. Place the collected blood in a 

 glass tube plus an anticoagulant and allow it to stand for several hours 

 or use mild centrifugation. The cellular elements will settle out and 

 leave a straw-colored solution on top. The leukocytes are not as dense 

 as the erythrocytes and will settle out last to remain as the so called 

 "buffy coat" between the red cell layer and the plasma on top. Smear 

 this "buffy coat" with its concentration of leukocytes on the slide and 

 fix. 



Klinger and Ludwig (1957) 



fixation: 95% alcohol for smears and thin embryonic membranes: 30 

 minutes to 4 hours. Tissue blocks in Davidson's fixative (see below): 

 3-24 hours and transfer to 95% alcohol. If material was fixed in some 

 other fixative, transfer immediately to 95%, alcohol, but do not leave 

 in first fixative for longer than 24 hours for good results. 



