SECTION TWO 



17. Start the dehydration by rinsing briefly in 50%, 70%, 

 80% and 90% alcohol. 



18. Rinse well in 95% alcohol. 



19. Finish the dehydration by immersing for four minutes in 

 each of two or three changes of absolute alcohol. 



20. Dry carefully with filter paper. 



21. Clear by immersion for four minutes each in two changes 

 of xylol. 



22. Mount in Canada in xylol or D.P.X. 



Results: 



Negri bodies: deep red, with the internal structure clearly 

 delineated. Some of the Negri bodies showed inclusions like 

 small granules encircled by a lighter halo, with basophilic granula- 

 tions inside. Others had a dispersed fine granulation inside, 

 intensified by their strong acidophilia and a large inclusion, also 

 acidophilic, with a slightly basophilic centre. The Negri bodies, 

 by reason of their characteristic internal structures, were clearly 

 differentiated from erythrocytes. The nucleoli were shown as 

 basophilic and could not possibly be mistaken for Negri bodies. 



Notes: 



(a) Of the three eosin solutions, the authors consider that 

 No. 3 (i.e. solution H) gives the best results. 



{b) The authors state that in view of their experience in using 

 an alcoholic solution of acid eosin of (Schleicher, 1953) and another 

 type (Kopsch, 1949-50) they are of opinion that the eosin used 

 in their technique (Massignani and Malferrari, 1961) does not 

 act solely as an acid eosin, but rather as a dye-mordant compound 

 in which pH is not important. 



(c) Copies of the original paper are obtainable from the authors, 

 in English, Portuguese, Italian and French, as well as colour 

 pictures and slides. Readers should refer to the original paper 

 for further information. 



Reference: Massignani, A. M. and Malferrari, R. C1961). 



