854 Histochemistry (chap. 21) 



use. Keep in bath during procedure. Other reagents at room tempera- 

 ture. 



1. Deparaffinize slides and run down to 70% alcohol. 



2. Place in a-naphthol: 15-30 minutes. 



3. Transfer (no wash) into hypochlorite-potassium hydroxide: 1.5 

 minutes. 



4. Transfer quickly into urea-potassium hydroxide, 2 changes. 

 First change: 10 seconds, agitate slides. 



Second change: 2 minutes. 



5. Transfer into absolute tertiary butyl alcohol, 2 changes. 

 First change: 10 seconds, agitate. 



Second change: 3.5 minutes. 



6. Transfer into xylene, 3 changes. 

 First change: 10 seconds, agitate. 

 Second change: 1-2 minutes. 

 Third change: 2-3 minutes. 



7. Drain off xylene, but do not dry. Mount in mineral oil. 



results: 



arsfinine — orans;e to red. Good for about 6 months. 



comments: 



Make all transfers as quickly as possible without allowing slides to 

 drain. Turn jars so slides do not hit lugs on sides. 



The first changes in steps 4, 5, and 6 are intended to wash off the 

 previous fluid, and the slides should be moved gently back and forth 

 to facilitate the process. This may be done in the second changes, but 

 more slowly. Timing must be accurate; a stop watch is advisable. 



Change solutions frequently; the reagents must be fresh. Carry not 

 more than 5 slides through the a-naphthol and hypochlorite-potas- 

 sium hydroxide and not more than 30 through any of the other solu- 

 tions. 



Solidification of the tertiary butyl alcohol may be caused by the 

 transfer of cold slides into it, so it is advisable to warm it to 30-40 °C 

 before use. Change the alcohol frequently. Carry about 10 slides 

 through 1 change; the first solution may then be discarded and re- 

 placed by the second change. The latter is then replaced by a fresh 

 solution. 



Before mounting, drain off the xylene thoroughly, but do not dry. 

 Apply several drops of mineral oil, and add cover glass. Draw off ex- 

 cess mineral oil with filter paper. (The excess oil is desirable because 



