STAINING, PRACTICAL AND THEORETICAL 



13. Drain and draw off excess water by means of a filter paper 

 applied carefully to the edges of the section, but do not allow the 

 preparation to dry completely. 



14. Dehydrate with absolute alcohol only. 



15. Clear in xylol. 



16. Mount in D.P.X., Cristalite or Clearmount. 



Results: 



Nuclei: dark brown to black. Collagen fibres: bright red. 

 Erythrocytes, muscle, epithelia and other tissues : yellow. 



Note: Van Gieson stain fades if mounted in Canada balsam, but 

 the degree of fading can be reduced by the use of D.P.X., or 

 Cristalite or Clearmount. 



References : 



Carleton, H. M. & Leach, E. H. (1947), pp. 128-129. 

 Van Gieson, J. (1889). 



HEXAZONIUM PARAROSANILINE - 

 ALPHANAPHTHYL PHOSPHATE 



For the localization of acid phosphatase 



Solutions required: 



A. Formalin (formaldehyde 40%) . . 10 ml. 

 Calcium chloride . . . . i gm. 

 Distilled water . . . . . . 90 ml. 



B. Sodium acetate, trihydrate, A.R. 9714 gm. 

 Sodium diethyl barbiturate (sol- 

 uble barbitone) . . . . 14714 gm. 



Boil about 500-600 ml. of distilled water to drive 

 off the dissolved CO2; then cool. 



Dissolve the two solids in about 450 ml. of the 

 C02-free distilled water; then make up the volume 

 to 500 ml. exactly, with C02-free water. 



C. Pararosaniline chloride . . . . 4 gm. 

 Hydrochloric acid, 2N . . . . 100 ml. 



244 



