VERHOEFF VANGEISON STAIN

       VERHOEFF VANGEISON STAIN

Principle

The disulphide bridges in the elastic fibers are converted to anionic sulphonic acid derivatives by treatment with iodine. These derivatives are strongly basophilic and capable of selective reaction with the basic dye (Haematoxylin).

Solutions

Solution A

Haematoxylin              -           5gm

Absolute alcohol          -           100ml

Solution B

Ferric Chloride            -           10gM

Distilled water             -           100 ml

Solution C

Iodine                           –         1 gm

Potassium iodide        -           2 gm

Distilled water                -           100 ml

Working Solution

Solution A         -           1ml

Solution B         -           0,4 ml

Solution C         -           0.4ml

Ferric chloride .2%

Ferric Chloride               -           2gm

Distilled water                -           l00 ml

Vangeison Solution

Saturated aqueous picric acid              –       100 ml

1%Acid fuchsin                                   –        10 ml

Procedure

1.      Deparaffinise the Section in Xylene for 10 mnts

2.      Bring the section to water through descending series of alcohol

3.      Treat the section with the solution mixture for 20-30 mts

4.      Wash it in water..

5.      Differentiate it in 2% ferric chloride until elastic tissue fibers appear black grey background. Differentiation should be checked under microscope .

6.      Keep it in running tap water for 5-10 mts

7.      Counterstain with Vangeisonsolution for 2 mts

8.      Blot it with filter paper, clear and Mount

Result

Elastic Fibre                 -           Black

Collagen                       -           Red

Muscle                         -           Yellow