STAINING, PRACTICAL AND THEORETICAL 



PHLOXIN - TARTRAZINE 



A general histological stain and for the demonstration of 



inclusion bodies 



Solutions required: 



A. Haemalum (Mayer). 



B. Calcium chloride 0-5% aqueous. 100 ml. 

 Phloxine . . . . . . . . 0-5 gm. 



C. Tartrazine, saturated in cellosolve. 



Technique: 



1 . Stain for five to ten minutes with the haemalum, examining 

 under the microscope at intervals, until the desired depth of 

 staining has been attained. 



2. Wash and blue in tap water or in saturated aqueous lithium 

 carbonate. 



3. Stain in the phloxine solution for half an hour. 



4. Rinse quickly in water. 



5. Drain off excess water and replace with tartrazine solution 

 (Solution C, as above), using a dropping bottle to control the 

 differentiation. 



Note: The tartrazine replaces the phloxine from collagen. As 

 tartrazine is readily soluble in water, slight overstaining is re- 

 commended before dehydration. 



6. Rinse in 60% alcohol followed by 95% alcohol. 



7. Dehydrate with absolute alcohol. 



8. Clear in xylol and mount. 



Results: 



Kurloff bodies in guinea pig's lung are well shown. Inclusion 

 bodies of a number of virus-containing tissues show retention of 

 phloxine. 



Notes: (a) Fixatives containing mercuric chloride give the 

 best results. 



(b) The author (Landrum) claims that this technique, in which 

 use is made of a stable phloxine solution, gives a brilliant demon- 

 stration of certain inclusion bodies and is superior to Masson's 

 erythrosin-saffron, which deteriorates fairly rapidly. 



Reference: Lendrum, A. C. (i947)- 



368 



