3G2 Special Procedures I (chap. 22) 



Add 2 ml. of "Tween 80" to 1000 ml. of stock. 



Working solution: 



acridine orange stock solution 1 part 



Mcllvaine's buffer 9 parts 



procedure: 



1. Hydrate slides to water, 5 dips in each of 80%, 70% and 50% al- 

 cohol. 



2. Dip 4 times in acetic acid, 1% (1 ml./99 ml. water). 



3. Rinse in distilled water: 2 minutes. 



4. Transfer to Mcllvaine's buffer: 3 minutes. 



5. Stain in buffered acridine orange: 3 minutes. 



6. Differentiate in Mcllvaine's btiffer: 4 miniues. 



7. Wipe excess material from ends of slides, gently blot with coarse 

 blotting paper. 



8. Apply cover glass with buffer. Let stand 2 minutes before examin- 

 ing. 



Can be restained, as above, with Papanicolaou. After this, can be de- 

 colorized and restained with acridine oran2:e. 



'o^ 



Sex Chromatin 



Greenblatt and Manautou (1957) 



fixation: peripheral blood smears in methyl alcohol: 1-2 minutes. 

 See Comments below for "buffy coat" suggestion. 

 Oral mucosal and vaginal smears in absolute alcohol-ether (1:1): 2-24 

 hours. 



solution: 

 Pinacyanole: 



pinacyanole* 0.5 gm. 



70% ethyl or methyl alcohol 100.0 ml. 



Wright's buffer: 



monobasic potassium phosphate 6.63 gm. 



dibasic sodium phosphate 3.20 gm. 



distilled water 1000.0 ml. 



PROCEDURES 



1. Cover with pinacyanole; blood smears: 30 seconds; oral mucosal 

 and vaginal smears: 45 seconds. 



* Eastman Kodak Co. 



