SECTION TWO 



SAFRANIN - WATER BLUE (Unna) 

 For collagen fibres 



Solutions required: 



A. Safranin O i% aqueous. 



B. Water Blue i% aqueous. . . . lo ml. 

 Tannic acid 33% aqueous . . 10 ml. 



This solution must be freshly prepared. 



Technique: 



1. Tissues should be fixed in 1% aqueous picric acid or in 

 absolute alcohol, and Celloidin sections should be employed. 



2. Stain for ten minutes in the safranin solution; then wash 

 thoroughly in water. 



3. Stain for ten to fifteen minutes in Solution B. 



4. Wash thoroughly in distilled water. 



5. Clear in Bergamot oil; then mount. 



Results: 



Collagen fibres are stained blue, while nuclei are red 



SCARLET R - ETHYLENE GLYCOL 



An improved technique for staining fat, etc, in animal tissues, 

 its chief advantages being: (a) Excellent differentiation without 

 loss of stain out of the lipid particles, {b) A stable solution which 

 does not dissolve lipid materials, {c) Sections are not shrunken 

 but remain pliable, {d) More intense staining of fat. 



Solutions required: 



A. Ethylene glycol, pure, anhydrous. 



B. Scarlet R . . . . ^ . . . . i gm. 

 Ethylene Glycol, pure, anhydrous. . 100 ml. 



Heat the ethylene glycol to 100-110° C. on 

 a hot plate or in an oven, or over the bunsen 

 flame, taking care that it does not catch fire; 

 then add the stain and stir until all or most of 

 it is dissolved. Filter when cold. 



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