Chapter VI: The Paraffin Method 51 



If the operator is accustomed to manipulate the fine adjustment 

 of his microscope with the left hand and the slide on the stage 

 with the right, it is best to have the label on the right end of 

 the slide; if not, then on the left. 



NOTE. Prepare four slides each of the other objects which 

 have been imbedded. Stain and mount one of each kind as you 

 did the intestine, and also one of each kind in the same way, 

 only substitute borax-carmine for the hematoxylin. The borax- 

 carmine may require 6 to 12 hours for staining. Preserve the 

 others for double staining. 



As time permits prepare and section the other tissues which 

 were fixed in alcohol and Gilson. After you have had the 

 preliminary practice in double staining, stain and mount these as 

 you prefer. 



II. DOUBLE STAINING IN HEMATOXYLIN AND EOSIN 



1. Proceed according to the regular schedule with one each of 

 the slides reserved above, and stain in Delafield's hematoxylin. 



2. Wash the sections in water, and proceed farther according 

 to the regular schedule to 95 per cent, alcohol. 



3. Transfer the slide to the eosin stain for 10 to 30 seconds, 

 and after rinsing again in 95 per cent, alcohol, place it in absolute 

 alcohol. 



4. Clear in carbol-xylol and mount in balsam. 



NOTE. The sections should show both the blue stain (in nuclei) and 

 the red stain (in cytoplasm) when examined under the microscope. If 

 either is too dense or too light, make a note of the fact and vary the time 

 accordingly when staining other sections by this method. 



III. DOUBLE STAINING IN CARMINE AND LYONS BLUE 



1. Pass the remaining reserved slides through xylol and the 

 alcohols, descending to 35 per cent, alcohol. 



2. Stain in borax-carmine for from 30 minutes to several hours, 

 until the sections are well colored. 



3. Rinse in water or 35 per cent, alcohol, and pass the sections 

 up through the alcohols to 95 per cent. If the sections are 

 deeply stained, however, remove the excess of stain with acid 



