STAINING, PRACTICAL AND THEORETICAL 



E. Reducer {after A. Palmgren, 1951) 

 Solution D . . . . . . . . 10 ml. 



Nitric Acid, aqueous . . . . i ml. 



Absolute alcohol . . . . • • 56 ml. 



Distilled water . . . . . . 44 ml. 



F. Gold chloride (brown or yellow), 



0-5% aqueous 

 *G. Oxalic acid, 1% aqueous 

 H. Sodium thiosulphate, 5% aqueous 

 I. Copper sulphate (CUSO4, 5H2O), 



20% aqueous 

 J. Silver nitrate, 7% aqueous 

 K. Bodian's developer {Bodian, 1936) 



Hydroquinone . . . . . . i gm. 



Sodium sulphite (Na2S03) . . . . 5 gm. 



Distilled water . . . . . . 100 ml. 



L. Oxalic acid, 2% aqueous 



*Note: Solutions B and G can be prepared by diluting 

 solutions I and L respectively. 



Technique I — Staining in toto 



Technique: 



1. Fix in solution A for twenty-four to forty-eight hours. 



2. Without washing, transfer to 70% alcohol for three to ten 

 days. 



3. Wash for about an hour in distilled water. 



4. Immerse for three hours in 10% copper sulphate (solution 

 B) at 50° C. 



5. Without washing, transfer to 1% silver nitrate (solution C) 

 for one to one and a half hours at 50° C. 



6. Wash for one minute in each of three lots of distilled water. 



7. Reduce by immersing in solution E at 40-45° C. for one to 

 five minutes. 



8. Wash well in distilled water. 



9. Tone in gold chloride 0-5% (solution F) for three to five 

 minutes, until the nervous system becomes visible. 



10. Wash in distilled water. 



11. Reduce in 1% oxalic acid (solution G) for one to three 

 minutes, until the nervous system is darkly stained. 



418 



