Staining Paraffin Sections 67 



Staining Chart II. The same slide can be repeatedly destained or 

 restained in the counterstain until exactly the desired effect is 

 obtained. The hematoxylin is not affected during this manijjulation. 



STAINING CHART II 



Hemalum With "General" Counterstain 

 Pre-Staining Operations and Intervals as in Chart I 



Hemalum 

 to correct 

 intensity 



i 

 30% 



alcohol 



i 



50% 



alcohol 



i 

 70% 



alcohol 



i 

 95% 



alcohol 



erythrosin 



(see text) > 



Other common counterstains used with the above hematoxylins, 

 are orange G, gold orange, eosin, fast green, and light green. The 

 underlying principle for applying other counterstains is the same as 

 for erythrosin. Counterstains may also be dissolved in clove oil and 

 applied after the last dehydrating step, omitting carbol-xylene because 

 clove oil is an excellent clearing agent. Counterstains may also be 

 dissolved in water, 50% to absolute alcohol, or Cellosolve and 

 introduced into the series at the corresponding point of dehydration. 



HEMALUM AND SAFRANIN 



After acceptable results have been obtained with the foregoing 

 single stain and the double stain, undertake the mastery of a double 

 stain having two selective components. One component of the next 

 double stain to be discussed is a self-mordanting hematoxylin; the 

 second component is safranin, which is highly selective for chromo- 

 somes, lignin, cutin, and in some cases for hemicellulose. An 

 important feature of this combination is that the hemalum is applied 

 to the desired intensity and remains fixed throughout subsequent 

 processing, whereas the safranin is applied until the material is 

 strongly overstained and then differentialh destained. 



