COPPER STAN-RUBEANIC ACID METHOD

       COPPER STAN-RUBEANIC ACID METHOD



Solutions

1) 0.1% Rubeanic acid in absolute alcohol               -        5ml
          10% Aqueous Sodium acetate                       -        100ml
                  
                             Freshly prepared prior to use
2) 1%Eosin
                   Eosin                     -        1 gm
                   Distilled water         -        100ml
                       
                        A few crystals of thymol are added.

Procedure
1)    Deparaffinise the section in xylene for 10mts.
2)    Bring the section to water through descending series of alcohol.
3)    The staining is done along with a known positive control.
4)    Rinse the section in distilled water.
5)    Place sections in a coplin jar filled with rubeanic acetate solution overnight at 37°C. 6) Wash in 70% alcohol.
6)    Rinse briefly in distilled water.
7)    Drain sections and blot dry.
8)    Counterstain with eosin for 30secs.
9)      Wash in water, dehydrate, clear and mount.


Result

                                    Copper                        -                       Greenish black
                                     Nuclei                         -                      Pink.


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