Journal of 



Applied Microscopy 



and 



Laboratory Methods 



Volume V. FEBRUARY, 1902. Number 2. 



Modification of Eosin and Mettiylen Blue Contrast- 

 staining, with Technique. 



Eosin and methylen blue contrast-stains have long been recognized as 

 extremely valuable in histological and pathological work, but many workers have 

 been deterred from their use by the difficulty of the technique, the lack of 

 uniformity in the results, and the comparative instability of the stain. 



The best method of using these stains at the present time is that employed by 

 Councilman and Mallory, but however well the stain has been perfected, there 

 have been many difficulties. The slight modifications employed by the author 

 consist in the addition of more potassium carbonate, making a stronger alkaline 

 methylen blue solution, and using a weaker solution of eosin. These modifica- 

 tions together with the definite instructions regarding technique enable one to 

 carry through a large number of sections in a comparatively short time with 

 uniformly more excellent results than have hitherto been obtained. The method 

 also seems to increase the permanency of the stains. 



The entire technique, including the fixing, hardening, imbedding, and stain- 

 ing of tissues, is given in detail : 



1. Harden pieces of tissue from 3 to 5 mm. in thickness in Zenker's fluid ^ for 

 twenty-four hours. 



2. Wash twenty-four hours in running water. 



3. Trim specimen to maximum size of 5x15x15 mm. and dehydrate in 95 

 jer cent, alcohol for twenty-four hours. 



4. Absolute alcohol twenty-four hours. 



5. Chloroform twenty-four hours. 



6. Chloroform and paraffin twenty-four hours. 



7. Paraffin at 120° F. four hours. 



8. Imbed and section specimen. Sections of internal organs should be cut 

 8 to 10 yu ; central nervous tissue, 5 to 8 yu. 



1. Zenker's fluid is the only satisfactory hardening and fixing reagent for this method, although 

 corrosive sublimate, Mueller's fluid, or Flemming's solution can be used. 



(1637) 



