SECTION TWO 



PINACYANOL - NEUTRAL RED 



For supra-vital staining of blood 



Note: Pinacyanol is superior to janus green in that it does not 

 fade : it is very selective for mitochondria, and it does not inhibit 

 the effect of neutral red. It has the disadvantage, however, of 

 being extremely costly. 



Solutions required: 

 Stock solutions: 



A. Pinacyanol o-i% in absolute alcohol. 



B. Neutral Red o-i% in absolute alcohol. 



Staining solutions: 



C. for mitochondria only 



Solution A . . . . . . . . I ml. 



Absolute alcohol . . . . . . 20 ml. 



D. for mitochondria, nuclei and other cell granules 

 Solution A . . . . . . . . I ml. 



Solution B . . . . . . . . 2 ml. 



Absolute alcohol . . . . . . 20 ml. 



Notes: 



(a) Solutions C and D deteriorate after a few hours and should, 

 therefore, be prepared only as and when they are required for 

 immediate use. 



(b) The proportions and dye concentrations of either of these 

 two solutions may be varied to suit particular specimens. 



Technique: 



1. Scrupulously clean dry slides are flooded with solution C 

 or D, whichever is required. 



2. Drain and leave to dry, then use as follows (within a few hours 

 at latest in the case of solution D as the dried dye combination 

 does not keep well). 



3. Place a drop of blood on each slide. 



4. Cover with scrupulously clean coverslip and allow the blood 

 to spread. 



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