Charcot-Leyden Crystal Protein Staining on Stool Samples 12/11/06 mkm

12/11/06 MKM

1. 
   Fix 20 ug of stool in 500 ul of PVA for 20 min.
   
2. 
   Use 20 ul of fixed stool on slide for smear.
   
3. 
   Dry overnite
   
4. 
   70% EtOH + iodine (until tea colored) 5-10 min. Removes mercuric chloride
   

5. 
   70 % EtOH 5 min. Removal of iodine & hydra.
   
6. 
   70% EtOH 3 min. Wash & hydration
   
7. 
   Trichrome (Wheatley’s) stain 10 min Staining
   
8. 
   90% Alcohol Acidified dip 1-3 sec Destaining
   

9. 
   Abs EtOH dip x 3 Stops staining
   
10. 
    Abs EtOH 3 min. Dehydration
    
11. 
    Abs EtOH 3 min. Dehydration
    
12. 
    Xylene 5-10 min. Removal of Alcohol
    
13. 
    Xylene 5-10 min. Removal of Alcohol
    
14. 
    Mount and examine under oil.