STAINING, PRACTICAL AND THEORETICAL 



Distilled water . . . . . . 50 ml. 



Note: This solution must be freshly prepared 

 immediately before use. 



B. Phosphate buffer solution pH 5-0 



Note: This can be conveniently prepared merely 

 by dissolving i Michrome buffer tablet pH 5-0 in 

 100 ml. of distilled water. 



C. Acridine red . . . . . . i gm. 



Distilled water . . . . . . 100 ml. 



Technique: 



1. Fix pieces of tissue in formol alcohol or formol saline. 



2. Dehydrate through the usual graded alcohols. 



3. Clear in xylol or cedarwood oil. 



4. Embed in paraffin wax by the usual procedure. 



5. Fix sections to slides and dewax as usual. 

 5. Carry through graded alcohols to water. 



7. Prepare a mixture of equal volumes of solutions A and B 

 sufficient to fill a Coplin jar (about 35 ml.). 



Note: This mixture deteriorates after a few hours. 



8. Immerse the slides in the stain in the Coplin jar for thirty 

 minutes at 37° C. in the incubator. The intensity of the staining 

 is poor at temperatures below 37° C. 



9. Wash in distilled water. 



10. Differentiate by direct immersion in absolute alcohol for 

 thirty minutes. 



11. Wash in distilled water. 



12. Counterstain with acridine red (solution C) for five minutes. 



13. Wash in distilled water. 



14. Differentiate rapidly in absolute alcohol. 



15. Mount in Clearmount: 



or 



16. Clear in xylol. 



17. Mount in D.P.X. or Emexel or Clearmount. 



Results: 



Sites of beryllium deposits are indicated by a clear apple-green 

 colorization against a red background. 



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