SECTION TWO 



C. Acid Alizarin Blue SWR. . . . 5 gm. 

 Aluminium sulphate 10% aqueous 100 ml. 



Boil for ten minutes. Cool and filter. Make up the 

 volume to 100 ml. with distilled water and add five 

 or six drops of formalin. 



D. Phosphomolybdic acid 5%. 



E. Alizarin viridin . . . . . . 0-2 gm. 



Buffer solution pH 5-8 . . 100 ml. 



Technique: 



1. Fix tissues in 10% formalin and embed in paraffin wax in 

 the usual manner. 



2. Fix sections to slides and take down to distilled water as 

 usual. 



3. Stain nuclei intensely by immersing the slides in the gallo- 

 cyanin solution in a staining jar, examining the preparations under 

 the microscope at intervals over a period of twenty-four hours, to 

 ascertain the depth of staining. 



4. Wash with two changes of distilled water. 



5. Stain elastic fibres in the orcein solution for ten minutes to 

 half an hour in a grooved, covered staining jar. 



6. Wash well with distilled water. 



7. Stain muscle in the acid alizarin blue solution for seven 

 minutes. 



8. Wash with distilled water. 



9. Differentiate in the phosphomolybdic acid solution for about 

 thirty minutes, controlling by examination under the micro- 

 scope at intervals. 



10. Wash with two changes of distilled water. 



11. Stain collagen in the alizarin viridin for seven minutes. 



12. Drain and blot thoroughly but carefully. 



13. Rinse with 96% alcohol; followed by carbol xylol. 



14. Wash well with two or three changes of xylol, and mount. 



Results: 



Nuclei: dark brown. Muscle and epithelium: pale violet. 

 Erythrocytes and elastic fibres are stained a rich brown, while 

 mucus, collagen are in varying shades of green ; myelin sheaths, 

 pink; and axis cylinders, dark blue. 



Reference: Buzaglio, J. H. (1934). 



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