STAINING, PRACTICAL AND THEORETICAL 



B. Safranin O, water soluble . . i gm. 



Aniline water . . . . . . 48 ml. 



Absolute alcohol . . . . 52 ml. 



Technique: 



1. Sections are mounted on slides and brought down to 90% 

 alcohol in the usual manner. (If the tissues have been fixed in a 

 fluid containing mercury, the mercurial deposit is removed by the 

 standard technique ; the sections are then taken from 70% to 90% 

 alcohol, then direct into the orcein stain.) 



2. Stain from twenty to sixty minutes with the orcein solution 

 in a stoppered staining jar. 



3. Rinse in acid alcohol until stain ceases to come out of the 

 preparation. 



4. Rinse in 70% alcohol; then in distilled water. 



5. Stain in aniline safranin (Solution B) for five minutes; then 

 rinse in water. 



6. Differentiate by dipping into 90% alcohol then examining 

 rapidly under the microscope. Repeat this process until the cell 

 nuclei are well brought out, stained clear bright red. 



7. Dehydrate by rinsing quickly in absolute alcohol; then clear 

 in xylol and mount in balsam or D.P.X. 



Results: 



Elastic fibres are stained dark to reddish brown; cell nuclei, 

 bright red; ground substance of hyaline cartilage, yellow. 



Reference: Carleton, H. M. & Leach, E. H. (1947), pp. 125-126. 



ORCEIN - GIEMSA STAIN 

 For syphilitic tissue, particularly dermatological specimens 



Solutions required: 

 A. Orcein (Unna-Tdnzer): 



B. 



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