SECTION TWO 



D. Sodium nitrite , . . . . . 4 gm 

 Distilled water . . . . . . 100 ml 



E. Sodium alphanaphthyl phosphate 0*02 gm 

 Distilled water . . . . . . 13 rnl 



Solution B . . . . . . 5 ml 



F. Solution C . . . . . . 0-8 ml. 



Solution D . . . . . . 0-8 ml. 



Drop the o-8 ml. of solution C into a test tube 

 and chill to o°-io° C. Add the o-8 ml. of solution D 

 a drop at a time, from a fine pipette, shaking the 

 tube after each addition, 



G. Add the contents of the test tube (solution F) to the 

 whole of solution E. 



Using a pH meter, adjust the pH of solution G, 

 which is the incubating medium, to 6*5 with N/1 

 NaOH (about 0-6 ml. will be required). 



Technique: 



1. Fix material for at least twenty-four hours, in solution A. 



Note: It is stated that a longer period of fixation (forty-eight to 

 seventy-two hours) is permissible, but this results in a small 

 decrease in activity. Fresh material is preferable, but autopsy 

 material of twenty-four hours gives fairly good results. 



2. Cut frozen sections and mount on albuminized slides that 

 have been dried at room temperature for one to two hours. 



3. Immerse in the freshly prepared solution G, at room 

 temperature, for five to thirty minutes, depending on the tissue. 



4. Rinse in distilled water, 



5. Dehydrate in alcohol. 



6. Clear in xylol. 



7. Mount in D.P.X., Clearmount or Exemel, or Canada 

 balsam in xylol. 



Results: 



Sites of acid phosphatase: deep brownish red, 



I 24s 



