MEDICAL AND BIOLOGICAL STAINING TECHNIQUES 



5. De-Stain for a few seconds in Solution C ; then wash in water. 



6. Immerse in Solution D for a few minutes; then rinse in 

 water. 



7. Stain for a few minutes in Solution E, then wash with water. 



8. Dehydrate very quickly with 95% and absolute alcohol; 

 clear in xylol and mount. 



Results: 



Bacteria, deep violet blue. Mitochondria and plastids, red. 



ACID RUBIN - AURANTIA - TOLUIDINE BLUE (KuU's 



Stain) 



For starch grains and mitochondria in plant tissues 



Solutions required: 



A. Acid rubin 1% aqueous. 



B. Aurantia 5% in 80% alcohol 



C. Tannic acid 2% aqueous. 



D. Toluidine blue 1% aqueous. 



Technique: 



1. Fix material in Regaud's fluid. 



2. Stain for five minutes in solution A heated to about 60 or 

 70° C. 



3. Differentiate with solution B, controlling by examination 

 under the microscope. 



4. Wash in water. 



5. Immerse in solution C for 20 minutes. 



6. Wash well in water. 



7. Stain for five minutes in solution D. 



8. Pour off excess stain and rinse with 70% alcohol. 



9. Differentiate in 90% alcohol. 



248 



