THE VENETIAN TURPENTINE METHOD 111 



much, wash it out in 95 per cent alcohol. The phloxine is not Hkely to over- 

 stain. 



5. Absolute alcohol, 5 or 6 seconds. 



6. Transfer quicklij to 10 per cent Venetian turpentine and proceed as in 

 the previous schedule. 



The surprising beauty of successful preparations will compensate 

 for whatever failures may occur. Nuclei and pyrenoids should show a 

 brilliant red, while the chromatophores and cytoplasm should be dark 

 blue. The cell walls should show a faint bluish color. 



Heidenhain's iron-alum haematoxylin and eosin. — Follow the 

 schedule for iron -haematoxylin until the glycerin has been washed out 

 in 95 per cent alcohol. Then stain for an hour in a solution of eosin in 

 95 per cent alcohol. Wash for a minute in 95 per cent alcohol, then a 

 minute in absolute alcohol, and then transfer to the 10 per cent Vene- 

 tian turpentine. 



Heidenhain's iron-alum haematoxylin and safranin. — Follow the 

 schedule for iron-haematoxylin until the glycerin has been washed out 

 in alcohol, and then add to the 95 per cent alcohol several drops of a 

 solution of safranin in 95 or 100 per cent alcohol and allow the stain to 

 act for 30 minutes or an hour. Then dehydrate in absolute alcohol and 

 transfer to 10 per cent Venetian turpentine. 



Other stains may be used. Aqueous stains should be used before 

 starting with 10 per cent glycerin. Alcoholic stains should be in strong 

 alcohol — about 90 per cent — and should be applied just after washing 

 out the glycerin. 



This method is equally good for filamentous fungi and also for the 

 prothallia of Equisetum and ferns, for delicate liverworts and mosses, 

 and similar objects. 



If you have a good turpentine, good stains, arid avoid moisture, the 

 Venetian turpentine method should not be difficult, and the results 

 with filamentous and unicellular forms and other small objects surpass 

 anything yet secured by other processes. 



